Sustained-release formulation for injection

ABSTRACT

The present invention relates to a composition comprising N-[4-[4-(1,2-benzisothiazol-3-yl)-1-piperazinyl]-(2R,3R)-2,3-tetramethylene-butyl]-(1′R,2′S,3′R,4′S)-2,3-bicyclo[2,2,1]heptanedicarboximide or a pharmaceutically acceptable acid addition salt thereof. In detail, the composition relates to a sustained release formulation for injection which maintains an effective blood level of the above-mentioned compound.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is a divisional of U.S. application Ser. No.15/043,235, filed Feb. 12, 2016, which is a Divisional of U.S.application Ser. No. 13/276,039, filed Oct. 18, 2011, now U.S. Pat. No.9,469,630, issued Oct. 18, 2016, which is based upon and claims thebenefits of priority to U.S. Provisional Application No. 61/394,069,filed Oct. 18, 2010. The entire contents of all of the aboveapplications are incorporated herein by reference.

TECHNICAL FIELD

The present invention relates to a composition comprisingN-[4-[4-(1,2-benzisothiazol-3-yl)-1-piperazinyl]-(2R,3R)-2,3-tetramethylene-butyl](1′R,2′S, 3′R,4′S)-2,3-bicyclo[2,2,1]heptanedicarboximide or apharmaceutically acceptable acid addition salt thereof (hereinafter,optionally referred to as “the present compound”). To be more specific,the present invention relates to a sterile suspension formulation whichcan maintain an effective blood level of the present compound as asustained release formulation for injection. Furthermore, the presentinvention relates to a process of the formulation comprising the presentcompound.

BACKGROUND ART

It is known thatN-[4-[4-(1,2-benzisothiazol-3-yl)-1-piperazinyl]-(2R,3R)-2,3-tetramethylene-butyl](1′R,2′S,3′R, 4′S)-2,3-bicyclo[2,2,1]heptanedicarboximide hydrochloride(hereinafter, optionally referred to as “Compound 1”) has psychotropicactivity and is useful as a therapeutic agent for diseases such asschizophrenia.

Patent Reference 1 describes that Compound 1 is usually administeredorally.

It is known that risperidone, olanzapine, aripiprazole, etc. have beenalready used in clinical practice as an agent for treating schizophreniaor other similar diseases. In order to promote patient compliance andreduce the relapse rate of schizophrenia, these medicaments for treatingschizophrenia are desired to be modified to exert a long-term effectthrough a single administration.

A depot formulation for intramuscular injection comprising an injectablesuspension of a medicament is generally known as one of administrationmeans for sustained-release of a medicament. An injectable suspension isin a heterogeneous system comprising a solid phase dispersed in anaqueous or nonaqueous liquid phase, which requires sterile, stable,resuspendable, syringeable, injectable, isotonic, and nonirritating. Inorder to develop such depot formulation, in particular, it is necessaryat least that such formulation is an aqueous one and additionally hassatisfactory properties for an injectable suspension such as uniformityof dispersed particles of an active ingredient, sedimentation propertyof dispersed particles after still standing, re-suspensibility, andpermeability in a needle (hereinafter, optionally referred to as“suspensibility”). In order to satisfy these requirements, for example,Patent Reference 2 discloses a composition comprising microparticlescomprising a polymeric binder, and an aqueous injectable vehicle. Thecomposition has a constructional property wherein the microparticleshave a mass median diameter of at least about 10 μm, and the vehiclecomprises a viscosity enhancing agent.

However, many of medicaments for treating schizophrenia or other similardiseases are hardly water-soluble, thus it is not easy to develop suchformulation thereof which can make these requirements satisfied.

For example, a formulation comprising aripiprazole makes suchrequirements satisfied by reducing the mean particle size of thecompound and adding a specific suspending agent such ascarboxymethylcellulose thereto (Patent Reference 3). And, a formulationcomprising olanzapine makes such requirements satisfied by furtherreducing the size of the compound to nanoparticles and using a surfacestabilizer such as polysorbate (Patent Reference 4). As just described,in order to prepare a suspension comprising such hardly water-solublecompound, it is necessary to select an adaptable means in response tothe property of the active ingredient after studying various means tomeet the requirements.

To prepare a formulation for injection, some extra processes such assterilization and dust free in a clean room are also required. Inparticular, a process under aseptic atmosphere is generally accompaniedwith some complicated and troublesome steps andmanufacturing-operations. In addition, to comply with the predeterminedlevel for assurance of aseptic preparation, some strict controls such assterilization of apparatuses and devices, education and training foroperators, and control of the number of environmental microbials andmicroparticles in an aseptic plant are also required.

PRIOR ART Patent Reference

[Patent Reference 1] JP Patent No. 2800953

[Patent Reference 2] JP 2003-534366 T

[Patent Reference 3] JP 2007-509148 T

[Patent Reference 4] JP 2008-520581 T

DISCLOSURE OF INVENTION Problem to be Solved by the Invention

Taking these conventional/current techniques into consideration, thepresent inventors attempted to improve the drug delivery property ofCompound 1 as a medicament for treating schizophrenia or other similardiseases and tried finding an injectable suspension formulation whichwould satisfy the desired suspensibility. Compound 1 has a lowsolubility in water, and exhibits poor water solubility andhydrophobicity. This means that it is technically difficult to formulatethe compound as an injectable product. To be more specific, whenCompound 1 is dispersed in water, the particles thereof tend to come upto the surface or adhere to the container wall, and thus it wasdifficult to satisfy the suspensibility as an injectable suspensionproduct. As a general technique to prepare an injectable suspensionproduct with a sustained release property, Patent Reference 4 teachesthat the releasing time can be adjusted by optimally controlling theparticle size and it is preferred to keep the mean particle size of anactive ingredient as small as possible. As for Compound 1, however, whenthe mean particle size thereof was too small, the sustained release fora long period could not be attained; whereas when the particles ofbigger mean particle size were used so that the sustained release for along period could be attained, then an injection needle was clogged withthe particles, thus it was found that such bigger particles wasdifficult for be injected.

A problem to be solved by the invention is to provide a formulation thatcan maintain an effective blood level of the present compound for a longperiod (hereinafter, optionally referred to as “the presentformulation”). In addition, the invention is intended to provide aprocess to accurately control the particle size of the present compound(hereinafter, optionally referred to as “the present process”).Furthermore, the invention is intended to provide a sterile anddust-free suspension formulation which can be easily processed withoutusing complicated dosage forms and/or complicated processes.

Means to Solve the Problem

The present inventors have extensively studied to reach the above objectand have surprisingly found that the above-mentioned problems could besolved by using a specific mean particle size of the present compound, aspecific nonionic surfactant, and an active ingredient having anoptimized concentration. The prepared suspension formulation forinjection thereby exhibits a good suspensibility and is able to passthrough an injection needle easily. Based upon the new findings, thepresent invention has been completed.

The present inventions are as follows:

Term 1

A composition comprising the following (1) to (5),

(1)N-[4-[4-(1,2-benzisothiazol-3-yl)-1-piperazinyl]-(2R,3R)-2,3-tetramethylene-butyl]-(1′R,2′S,3′R,4′S)-2,3-bicyclo[2,2,1]heptanedicarboximideor a pharmaceutically acceptable acid addition salt thereof as an activeingredient,

(2) a surfactant comprising at least one ingredient selected from thegroup consisting of polysorbate 80, polysorbate 20, polyoxyethylenehydrogenated castor oil 50, polyoxyethylene hydrogenated castor oil 60,poloxamer 188 and polyoxyethylene castor oil,

(3) a buffer comprising at least one ingredient selected from the groupconsisting of sodium phosphate, disodium hydrogenphosphate, sodiumdihydrogenphosphate, potassium phosphate, dipotassium hydrogenphosphate,potassium dihydrogenphosphate, trometamol, sodium carbonate, sodiumbicarbonate, meglumine, arginine, triethanolamine and citric acid,

(4) an isotonic agent comprising at least one ingredient selected fromthe group consisting of sodium chloride and D-mannitol, and

(5) water for injection.

Term 2

The composition of Term 1 wherein the active ingredient is in a crystalform.

Term 3

The composition of Term 2 wherein the mean particle size of the crystalis about 4 μm-about 55 μm, about 4 μm-about 26 μm, about 4 μm-about 20μm, about 4 μm-about 18 μm, about 4 μm-about 16 μm, about 5 μm-about 55μm, about 5 μm-about 26 μm, about 5 μm-about 20 μm, about 5 μm-about 18μm, about 5 μm-about 16 μm, about 8 μm-about 55 μm, about 8 μm-about 20μm, about 10 μm-about 55 μm, about 10 μm-about 26 μm, about 10 μm-about20 μm, about 10 μm-about 18 μm, or about 10 μm-about 16 μm.

Term 4

The composition of Term 3 wherein the mean particle size of the crystalis about 5 μm-about 55 μm.

Term 5

The composition of Term 3 wherein the mean particle size of the crystalis about 5 μm-about 26 μm.

Term 6

The composition of Term 3 wherein the mean particle size of the crystalis about 5 μm-about 20 μm.

Term 7

The composition of Term 3 wherein the mean particle size of the crystalis about 5 μm-about 16 μm.

Term 8

The composition of Term 3 wherein the mean particle size of the crystalis about 10 μm-about 20 μm.

Term 9

The composition of Term 3 wherein the mean particle size of the crystalis about 10 μm-about 16 μm.

Term 10

The composition of any one of Terms 2 to 9 wherein the crystal is acubic crystal.

Term 11

The composition of Term 10 wherein the ratio between length and width ofthe cubic crystal is about 1:1, and the ratio between length and heightthereof is about 1:0.8 to about 1:1.2.

Term 12

The composition of any one of Term 2 to 11 wherein the active ingredientin crystal form is contained in about 5% (w/v) to about 60% (w/v) perthe whole of the composition.

Term 13

The composition of any one of Terms 1 to 12 wherein the surfactant ispolysorbate 80.

Term 14

The composition of any one of Terms 1 to 13 wherein the surfactant iscontained in about 0.005% (w/v) to about 2% (w/v) per the whole of thecomposition.

Term 15

The composition of any one of Terms 1 to 14 wherein the buffer is atleast one ingredient selected from the group consisting of sodiumphosphate, disodium hydrogenphosphate, sodium dihydrogenphosphate,potassium phosphate, dipotassium hydrogenphosphate, potassiumdihydrogenphosphate, trometamol, sodium carbonate, sodium bicarbonate,meglumine, arginine, triethanolamine and citric acid.

Term 16

The composition of Term 15 wherein the buffer is sodiumdihydrogenphosphate and/or disodium hydrogenphosphate.

Term 17

The composition of any one of Terms 1 to 16 wherein the buffer iscontained in about 0.01% (w/v) to about 2% (w/v) per the whole of thecomposition.

Term 18

The composition of any one of Terms 1 to 17 wherein the isotonic agentis sodium chloride.

Term 19

The composition of any one of Terms 1 to 18 wherein the isotonic agentis contained in about 0.1% (w/v) to about 10% (w/v) per the whole of thecomposition.

Term 20

A sustained release formulation for injection which comprises thecomposition according to any one of Terms 1 to 19.

Term 21

The formulation of Term 20 wherein the active ingredient is contained ina concentration of about 50 mg/mL to about 600 mg/mL.

Term 22

The formulation of Term 21 wherein the active ingredient is contained ina concentration of about 50 mg/mL to about 400 mg/mL.

Term 23

The formulation of Term 22 wherein the active ingredient is contained ina concentration of about 100 mg/mL to about 400 mg/mL.

Term 24

The formulation of Term 23 wherein the active ingredient is contained ina concentration of about 100 mg/mL to about 200 mg/mL.

Term 25

The formulation of Term 23 wherein the active ingredient is contained ina concentration of about 200 mg/mL to about 400 mg/mL.

Term 26

The formulation of any one of terms 20 to 25 wherein the surfactant iscontained in a concentration of about 0.05 mg/mL to about 20 mg/mL.

Term 27

The formulation of any one of Terms 20 to 26 wherein the buffer iscontained in a concentration of about 0.1 mg/mL to about 20 mg/mL.

Term 28

The formulation of any one of Terms 20 to 27 wherein the isotonic agentis contained in a concentration of about 1 mg/mL to about 100 mg/mL.

Term 29

The formulation of any one of Terms 20 to 28 which contains about 50 mgto about 1200 mg of the active ingredient per a container.

Term 30

The formulation of Term 29 which contains about 100 mg to about 800 mgof the active ingredient per a container.

Term 31

The formulation of any one of Terms 20 to 30 which can pass through aneedle of 18 to 23 gauges.

Term 32

The formulation of any one of Terms 20 to 31 wherein the sustainedrelease formulation for injection is a depot formulation for injection.

Term 33

A process of a sustained-release sterile formulation for injectioncomprising the following steps (1) to (5):

step (1): dissolving a mixture comprisingN-[4-[4-(1,2-benzisothiazol-3-yl)-1-piperazinyl]-(2R,3R)-2,3-tetra-methylene-butyl]-(1′R,2′S,3′R,4′S)-2,3-bicyclo[2,2,1]-heptanedicarboximideor a pharmaceutically acceptable acid addition salt thereof and asurfactant with an organic solvent or a mixture of an organic solventand water, and then sterilely-filtering the solution,

step (2): preparing a water solution comprising a surfactant and abuffer, sterilely filtering the water solution, and then sterilelyputting the water solution into a sealed vessel whose inside issterilized,

step (3): sterilely adding the sterile solution prepared in step (1)into the sealed vessel containing the water solution prepared in step(2),

step (4): collecting a crystal precipitated in step (3) through a filterin the sealed vessel, and

step (5): preparing a water solution comprising a surfactant, a buffer,an isotonic agent and water for injection, sterilely filtering the watersolution, and then sterilely putting the water solution into the sealedvessel containing the crystal in step (4) and mixing the water solutionand the crystal.

Term 34

A process of a sustained-release formulation for injection comprisingthe following steps (1) to (5):

step (1): dissolving a mixture comprisingN-[4-[4-(1,2-benzisothiazol-3-yl)-1-piperazinyl]-(2R,3R)-2,3-tetra-methylene-butyl]-(1′R,2′S,3′R,4′S)-2,3-bicyclo[2,2,1]-heptanedicarboximideor a pharmaceutically acceptable acid addition salt thereof and asurfactant with an organic solvent or a mixture of an organic solventand water to prepare a solution of the active ingredient,

step (2): preparing a water solution comprising a surfactant and abuffer (i.e. a crystallizing agent),

step (3): adding the solution of the active ingredient prepared in step(1) into the crystallizing agent in step (2),

step (4): collecting a crystal precipitated in step (3) through afilter, and

step (5): preparing a water solution comprising a surfactant, a buffer,an isotonic agent and water for injection (i.e. a vehicle solution), andthen mixing the water solution and the crystal of the active ingredientprepared in step (4) to prepare a suspension formulation.

Term 35

A process of a sustained-release formulation for injection comprisingthe following steps (1) to (5):

step (1): dissolving a mixture comprisingN-[4-[4-(1,2-benzisothiazol-3-yl)-1-piperazinyl]-(2R,3R)-2,3-tetra-methylene-butyl]-(1′R,2′S,3′R,4′S)-2,3-bicyclo[2,2,1]-heptanedicarboximideor a pharmaceutically acceptable acid addition salt thereof and asurfactant with an organic solvent or a mixture of an organic solventand water to prepare a solution of the active ingredient,

step (2): preparing a water solution comprising a surfactant and abuffer (i.e. a crystallizing agent), and then injecting the watersolution into a vessel for crystallization,

step (3): adding the solution of the active ingredient prepared in step(1) into the vessel for crystallization containing the crystallizingagent in step (2),

step (4): collecting a crystal precipitated in step (3) through a filterin the vessel for crystallization, and

step (5): preparing a water solution comprising a surfactant, a buffer,an isotonic agent and water for injection (i.e. a vehicle solution), andthen injecting the water solution into the crystal of the activeingredient prepared in step (4) and mixing the water solution and thecrystal to prepare a suspension formulation.

Term 36

The process of any one of Terms 33 to 35 further followed by thefollowing steps (6) to (8):

step (6): aseptically filling a formulation container with thesuspension formulation prepared in step (5) to prepare a filledsuspension formulation,

step (7): sterilizing the filled suspension formulation prepared in step(6) by a process of steaming under pressure, and

step (8): ultrasonicating the filled suspension formulation prepared instep (7).

Term 37

The process of Term 36 wherein the formulation container is a prefilledsyringe or a vial.

Term 38

The process of Term 36 or 37 wherein the temperature of the steamsterilization under pressure is 100 to 150° C.

Term 39

The process of any one of Terms 33 to 38, in step (3) the solution ofthe active ingredient is added to the crystallizing agent in the vesselfor crystallization (or the sealed vessel) while circulating thesolution (or the suspension solution) in the vessel for crystallization(or the sealed vessel) via a bypass pathway equipped with a pump whichcan pressure a solution.

Term 40

The process of any one of Term 39 wherein the pump in the bypass pathwayis any one pump selected from a roller pump (tube pump, hose pump), areciprocating pump (piston pump, plunger pump, diaphragm pump), and arotary pump (gear pump, vane pump, screw pump).

Term 41

The process of Term 39 wherein the pump in the bypass pathway is aroller pump.

Term 42

The process of any one of Terms 39 to 41 wherein the circulationvelocity of the bypass pathway is about 0.001 to about 2 parts byvolume/min per one part of the total volume of the crystallizing agentand the solution of the active ingredient.

Term 43

The process of any one of Terms 39 to 41 wherein the circulationvelocity of the bypass pathway is about 0.01 to about 1 parts byvolume/min per one part of the total volume of the crystallizing agentand the solution of the active ingredient.

Term 44

A process of a sustained-release formulation for injection comprisingthe following steps (1) to (5):

step (1): dissolving a mixture comprisingN-[4-[4-(1,2-benzisothiazol-3-yl)-1-piperazinyl]-(2R,3R)-2,3-tetra-methylene-butyl]-(1′R,2′S,3′R,4′S)-2,3-bicyclo[2,2,1]-heptanedicarboximideor a pharmaceutically acceptable acid addition salt thereof and asurfactant with an organic solvent or a mixture of an organic solventand water to prepare a solution of the active ingredient,

step (2): preparing a water solution comprising a surfactant and abuffer (i.e. a crystallizing agent) and injecting the water solutioninto a vessel for crystallization equipped with a stirring system,

step (3): adding the solution of the active ingredient to thecrystallizing agent in the vessel for crystallization while circulatingthe solution (or the suspension solution) in the vessel forcrystallization via a bypass pathway equipped with a pump which (i) isconnected to the vessel for crystallization, (ii) can make the solutionin the vessel for crystallization circulated, and (iii) can pressure thesolution,

step (4): collecting a crystal precipitated in the vessel forcrystallization from the solution (or the suspension solution) in step(3) through a filter, and

step (5): preparing a water solution comprising a surfactant, a buffer,an isotonic agent and water for injection (i.e. a vehicle solution), andthen injecting the water solution into the crystal of the activeingredient prepared in step (4) and mixing them to prepare a suspensionformulation.

Term 45

A process of a sustained-release formulation for injection comprisingthe following steps (4) to (8):

step (4): preparing a vehicle solution comprising a surfactant, abuffer, an isotonic agent and water for injection,

step (5): injecting the vehicle solution in step (4) into a crystal ofN-[4-[4-(1,2-benzisothiazol-3-yl)-1-piperazinyl]-(2R,3R)-2,3-tetramethylene-butyl]-(1′R,2′S,3′R,4′S)-2,3-bicyclo[2,2,1]heptanedicarboximideand mixing them to prepare a suspension formulation,

step (6): aseptically filling a formulation container with thesuspension formulation prepared in step (5) to prepare a filledsuspension formulation

step (7): sterilizing the filled suspension formulation prepared in step(6) by a process of steaming under pressure, and

step (8): ultrasonicating the filled suspension formulation prepared instep (7).

Term 46

The process of any one of Terms 33 to 45 wherein each the surfactant isindependently at least one ingredient selected from the group consistingof polysorbate 80, polysorbate 20, polyoxyethylene hydrogenated castoroil 50, polyoxyethylene hydrogenated castor oil 60, poloxamer 188,polyoxyethylene castor oil, benzalkonium chloride and sodium laurylsulfate.

Term 47

The process of any one of Terms 33 to 46 wherein the organic solvent isat least one solvent selected from the group consisting of 1-propanol,methanol, ethanol, 2-propanol, acetone, dimethylsulfoxide andN,N-dimethylacetamide.

Term 48 The process of any one of Terms 33 to 47 wherein the solventused in step (1) is a mixture of an organic solvent and water.

Term 49

The process of Term 48 wherein the mixture solvent is a water-containingalcohol.

Term 50

The process of Term 49 wherein the content of alcohol in thewater-containing alcohol is 40% to 90%.

Term 51

The process of Term 49 wherein the content of alcohol in thewater-containing alcohol is 50% to 90%.

Term 52

The process of any one of Terms 33 to 51 wherein the alcohol is ethanol.

Term 53

The process of any one of Terms 33 to 52 wherein each the buffer isindependently at least one ingredient selected from the group consistingof sodium dihydrogenphosphate, disodium hydrogenphosphate, potassiumdihydrogenphosphate, dipotassium hydrogenphosphate, sodium carbonate,triethanolamine, arginine and meglumine.

Term 54

The process of any one of Terms 33 to 53 wherein the isotonic agent issodium chloride and/or D-mannitol.

Term 55

A sustained release formulation for injection which is prepared throughthe process of any one of Terms 33 to 54.

Term 56

A crystal ofN-[4-[4-(1,2-benzisothiazol-3-yl)-1-piperazinyl]-(2R,3R)-2,3-tetramethylene-butyl]-(1′R,2′S,3′R,4′S)-2,3-bicyclo[2,2,1]heptanedicarboximidewhose mean particle size is in a range selected from the groupconsisting of about 4 μm-about 55 μm, about 4 μm-about 26 μm, about 4μm-about 20 μm, about 4 μm-about 18 μm, about 4 μm-about 16 μm, about 5μm-about 55 μm, about 5 μm-about 26 μm, about 5 μm-about 20 μm, about 5μm-about 18 μm, about 5 μm-about 16 μm, about 8 μm-about 55 μm, about 8μm-about 20 μm, about 10 μm-about 55 μm, about 10 μm-about 26 μm, about10 μm-about 20 μm, about 10 μm-about 18 μm, and about 10 μm-about 16 μm.

Term 57

The crystal of Term 56 wherein the mean particle size of the crystal isabout 5 μm-about 55 μm.

Term 58

The crystal of Term 56 wherein the mean particle size of the crystal isabout 5 μm-about 26 μm.

Term 59

The crystal of Term 56 wherein the mean particle size of the crystal isabout 5 μm-about 20 μm.

Term 60

The crystal of Term 56 wherein the mean particle size of the crystal isabout 5 μm-about 16 μm.

Term 61

The crystal of Term 56 wherein the mean particle size of the crystal isabout 10 μm-about 20 μm.

Term 62

The crystal of Term 56 wherein the mean particle size of the crystal isabout 10 μm-about 16 μm.

Term 63

A crystal ofN-[4-[4-(1,2-benzisothiazol-3-yl)-1-piperazinyl]-(2R,3R)-2,3-tetramethylene-butyl]-(1′R,2′S,3′R,4′S)-2,3-bicyclo[2,2,1]heptanedicarboximide(free form of Compound 1) which has a powder X-ray diffraction patternshowing diffraction angle 2θ (°) of 15.1±0.2, 15.5±0.2, 16.3±0.2,16.6±0.2, 18.0±0.2, and 20.0±0.2.

Term 64

A crystal ofN-[4-[4-(1,2-benzisothiazol-3-yl)-1-piperazinyl]-(2R,3R)-2,3-tetramethylene-butyl]-(1′R,2′S,3′R,4′S)-2,3-bicyclo[2,2,1]heptanedicarboximide(free form of Compound 1) which has a powder X-ray diffraction patternshowing diffraction angle 2θ (°) of 11.2±0.2, 15.1±0.2, 15.5±0.2,16.3±0.2, 16.6±0.2, 18.0±0.2, 19.1±0.2, 19.5±0.2, 20.0±0.2, 20.9±0.2,22.3±0.2, and 26.0±0.2.

Term 65

The crystal of any one of Terms 56 to 62 which has a powder X-raydiffraction pattern showing diffraction angle 2θ (°) of 15.1±0.2,15.5±0.2, 16.3±0.2, 16.6±0.2, 18.0±0.2, and 20.0±0.2.

Term 66

The crystal of any one of Terms 56 to 65 wherein the crystal is a cubiccrystal.

Term 67

The crystal of Term 66 which is a cubic crystal wherein the ratiobetween length and width of the cubic crystal is about 1:0.8 to about1:1.2, and the ratio between length and height thereof is about 1:0.1 toabout 1:3.

Term 68

A depot formulation for injection comprising the crystal of any one ofTerms 56 to 67.

Term 69

The depot formulation for injection of Term 68 wherein the crystal issuspended in a medium of the formulation.

Term 70

The depot formulation for injection of Term 69 wherein the medium iswater.

Term 71

The depot formulation for injection of any one of Terms 68 to 70 whichfurther comprises a surfactant, a buffer and an isotonic agent.

Term 72

A package containing a container which contains the sustained releaseformulation for injection comprising the crystal of any one of Terms 56to 67.

Term 73

A method for treating psychiatric disease which comprises administeringthe formulation of any one of Terms 20 to 32, 55, and 68 to 71.

Term 74

The method of Term 73 wherein the psychiatric disease is schizophrenia.

Term 75

The method of Term 73 wherein the psychiatric disease is bipolardisorder.

Term 76

The method of Term 73 wherein the psychiatric disease is depression.

Effect of the Invention

The present formulation can maintain an effective blood level of thepresent compound for at least 2 to 4 weeks or more by intramuscularadministration. Consequently, using the present formulation, it ispossible to decrease the frequency of administration compared with anoral formulation containing the present compound. The presentformulation is injectable (i.e. has a good needle passability) eventhough the present compound is suspended in the present formulation. Thepresent process provides the formulation to maintain an effective bloodlevel of the present compound for at least 2 to 4 weeks or more bysuspending the present compound in the formulation. In addition, thepresent process is useful in the point that the preparation is carriedout under a sequential sterile and/or dust-free processing, further viaa continuous formulating process.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a photomicrograph of the crystal ofN-[4-[4-(1,2-benzisothiazol-3-yl)-1-piperazinyl]-(2R,3R)-2,3-tetramethylene-butyl]-(1′R,2′S,3′R,4′S)-2,3-bicyclo[2,2,1]-heptanedicarboximide.

FIG. 2 is a graph showing a transition of the average concentration ofdrug in the plasma when the formulation prepared in Example 12 (200mg/mL) was administered to rats in a dose of 50 mg/kg.

FIG. 3 is a graph showing a transition of the average concentration ofdrug in the plasma when the formulation prepared in Example 13 (400mg/mL) was administered to rats in a dose of 100 mg/kg.

FIG. 4 is a photograph showing that the non-ultrasonicated formulationprepared in Example A2 contains macroaggregated particles.

FIG. 5 is a photograph showing that the ultrasonicated formulation (35kHz, 3 min) prepared in Example A2 contains no macroaggregatedparticles.

FIG. 6 is an X-ray diffraction in the sample before the steamsterilization under pressure in Example A3.

FIG. 7 is an X-ray diffraction in the sample after the steamsterilization under pressure (121° C., 20 min) and the ultrasonication(35 kHz, 3 min) in Example A3.

BEST MODE FOR CARRYING OUT THE INVENTION

The term “active ingredient” used herein isN-[4-[4-(1,2-benzisothiazol-3-yl)-1-piperazinyl]-(2R,3R)-2,3-tetra-methylene-butyl]-(1′R,2′S,3′R,4′S)-2,3-bicyclo[2,2,1]-heptanedicarboximideor a pharmaceutically acceptable acid addition salt thereof. Preferably,the term refers toN-[4-[4-(1,2-benzisothiazol-3-yl)-1-piperazinyl]-(2R,3R)-2,3-tetramethylene-butyl]-(1′R,2′S,3′R,4′S)-2,3-bicyclo[2,2,1]-heptanedicarboximideor a hydrochloride thereof. The concentration of the active ingredientused herein is within about 5% (w/v) to about 60% (w/v) per the whole ofthe formulation as a free form.

In addition, the concentration of the active ingredient in theformulation is preferably about 50 mg/mL to about 600 mg/mL as a freeform, and more preferably about 50 mg/mL to about 400 mg/mL, about 100mg/mL to about 400 mg/mL, about 100 mg/mL to about 200 mg/mL, or about200 mg/mL to about 400 mg/mL as a free form.

The “particle size” is controlled to maintain an effective blood levelof the active ingredient for at least two weeks, preferably for at leastfour weeks. The mean particle size of the “active ingredient” usedherein is in a range selected from the group consisting of about 4μm-about 55 μm, about 4 μm-about 26 μm, about 4 μm-about 20 μm, about 4μm-about 18 μm, about 4 μm-about 16 μm, about 5 μm-about 55 μm, about 5μm-about 26 μm, about 5 μm-about 20 μm, about 5 μm-about 18 μm, about 5μm-about 16 μm, about 8 μm-about 55 μm, about 8 μm-about 20 μm, about 10μm-about 55 μm, about 10 μm-about 26 μm, about 10 μm-about 20 μm, about10 μm-about 18 μm, and about 10 μm-about 16 μm. Preferably, the meanparticle size is about 5 μm-about 55 μm, about 5 μm-about 26 μm, about 5μm-about 20 μm, about 5 μm-about 16 μm, about 10 μm-about 20 μm, orabout 10 μm-about 16 μm. The term “mean particle size (also referred toas D50)” used herein is defined as a median diameter based on thevolume, which was obtained by measuring the particle size distributionthrough a wet process with a laser diffraction particle sizedistribution analyzer.

The term “pharmaceutically acceptable acid addition salt” used herein isdefined as an addition salt with an inorganic acid or an organic acid.The inorganic acid includes, for example, hydrochloric acid, hydrobromicacid, hydroiodic acid, and sulfuric acid. The organic acid includes, forexample, phosphoric acid, acetic acid, oxalic acid, citric acid, malicacid, tartaric acid, maleic acid, and fumaric acid. Preferably, the saltis hydrochloride.

The “surfactant” is used to improve the suspensibility of the crystal inthe present formulation, and is contained in a range of about 0.005 toabout 2% (w/v) per the whole of the formulation, preferably about 0.01to about 1% (w/v). In addition, the surfactant is contained in aconcentration of about 0.05 mg/mL to about 20 mg/mL. The surfactantincludes, for example, polysorbate 80, polysorbate 20, polyoxyethylenehydrogenated castor oil 50, polyoxyethylene hydrogenated castor oil 60,poloxamer 188, and polyoxyethylene castor oil. Preferably, thesurfactant is polysorbate 80. In addition, the present invention maycontain more than two surfactants.

The “buffer” is used in an appropriate amount to adjust the pH of theaqueous suspension in the present formulation to an injectable range,for example about pH 6 to about pH 8 and preferably about pH 7. Thebuffer is contained in a range of about 0.01 to about 2% (w/v) per thewhole of the formulation, and preferably about 0.05 to about 1% (w/v).In addition, the buffer is contained in a concentration of about 0.1mg/mL to about 20 mg/mL. The buffer includes, for example, sodiumphosphate, disodium hydrogenphosphate, sodium dihydrogenphosphate,potassium phosphate, dipotassium hydrogenphosphate, potassiumdihydrogenphosphate, trometamol, sodium carbonate, sodium bicarbonate,meglumine, arginine, triethanolamine, and citric acid. Preferably, thebuffer is sodium phosphate. In addition, the present invention maycontain more than two buffers.

The “isotonic agent” is used to adjust the osmotic pressure of thepresent formulation to an injectable range. The isotonic agent iscontained in a range of about 0.1 to 10% (w/v) per the whole of theformulation, preferably about 0.2 to about 5% (w/v). In addition, theisotonic agent is contained in a concentration of about 1 mg/mL to about100 mg/mL. The isotonic agent includes, for example, sodium chloride,and D-mannitol. Preferably, the isotonic agent is sodium chloride.

The term “water” used herein is defined as purified water, or the sameor a higher grade thereof; and such water needs to be sterilized afterdissolving various ingredients therein, or already-sterilized water(e.g. water for injection) is used in the process which is carried outunder sterile condition throughout the steps. In addition, the term“water for injection” used herein includes water sterilized through asterile filter etc. after dissolving substrates or reagents into theabove-mentioned “water” (as a starting material).

As shown in FIG. 1, the crystal ofN-[4-[4-(1,2-benzisothiazol-3-yl)-1-piperazinyl]-(2R,3R)-2,3-tetra-methylene-butyl]-(1′R,2′S,3′R,4′S)-2,3-bicyclo[2,2,1]-heptanedicarboximideor a pharmaceutically acceptable acid addition salt thereof ischaracterized by a cubic crystal form just after being prepared throughthe present process. And then, the crystal might be gradually roundedand transformed into a spherical form during storage. In addition, thecubic crystal used herein includes not only perfect cubic shapes butalso cuboidal shapes (e.g. shapes whose ratio between length and widthis about 1:0.8 to about 1:1.2, and the ratio between length and heightthereof is about 1:0.1 to about 1:3, etc.).

The “sustained release formulation for injection” used herein means aformulation which can maintain the blood level of the active ingredientfor at least two to four weeks after intramuscular injection.

The present formulation may contain other additive agents unless thereare any adverse effects on the present invention. The other additiveagents includes, for example, pH regulators such as hydrochloric acid,acetic acid, lactic acid, sulfuric acid, phosphoric acid,methanesulfonic acid, tartaric acid, and sodium hydroxide; soothingagents such as lidocaine hydrochloride, meprylcaine hydrochloride andprocaine hydrochloride; water-soluble polymers such ascarboxymethylcellulose sodium, hydroxypropylcellulose, hydroxypropylmethylcellulose, hydroxymethylcellulose, methylcellulose,polyvinylpyrrolidone, polyvinyl alcohol, polyethylene glycol 4000,carboxy vinyl polymer, polypeptide, polyamino acid, dextrin, gum arabic,sodium alginate, pullulan, sodium hyaluronate, sodium chondroitinsulfate, gelatin, collagen, agarose, xanthane gum, gellan gum, tamarindgum, guar gum, carrageenan, locust bean gum, karaya gum, tragacanth gum,tara gum, psyllium seed gum, ghatti gum, curdlan, and pectin; and otherpolymers such as sulfobutylether β-cyclodextrin, hydroxypropylβ-cyclodextrin, polylysine, polyglutamic acid, chitin, chitosan,polylactic acid, polyglycolic acid and sucrose acetate isobutylate.

The present formulation can be administered parenterally, in particular,intramuscularly or subcutaneously as a sustained release formulation forinjection, at a dose of 50 to 1200 mg (preferably 100 to 800 mg) of theactive ingredient. The present formulation can be administered once intwo to six weeks, preferably once in four to six weeks, and morepreferably once in four weeks.

The present formulation may contain 50 to 1200 mg of the activeingredient per a bottle, and preferably 100 to 800 mg thereof.

The present formulation can be administered intramuscularly orsubcutaneously with a prefilled syringe equipped with a needle which isfilled with the present formulation. Alternatively, the presentformulation can be administered through the following steps; i.e., froma vial filled with the present formulation, the content is transferredinto an injection syringe via a needle and then administeredintramuscularly or subcutaneously. Furthermore, the present formulationmay be put into a container such as a vial and then lyophilized to givea lyophilize formulation, or the crystal of the active ingredient in thepresent formulation may be isolated, dried, and then put into acontainer such as a vial to give a powder-filled formulation. Thelyophilize formulation or the powder-filled formulation can beadministered intramuscularly or subcutaneously as an injectablesuspension which is prepared by putting a suspending solution into thecontainer via a needle just before use. A needle of 18 gauge (internaldiameter thereof: 0.90 mm) to 23 gauge (internal diameter thereof: 0.33mm) may be used for the intramuscular or subcutaneous administration.Furthermore, the container filled with the formulation of the presentinvention can be set in a needleless syringe which can administer a drugsolution in the container by discharging the solution with a pressureprepared by a gas, a detonator, a spring, etc. packed in the syringedevice without using any needle to administer the drug solutionintramuscularly or subcutaneously. The formulation of the presentinvention includes a form of liquid such as a suspension which can beput in a formulation container, a form wherein the liquid such as asuspension is already put in a formulation container, and further a formwherein the formulation container containing the liquid is sterilizedand/or ultrasonicated.

The present formulation can be prepared according to, for example, thefollowing steps, but are not limited thereto.

Preparation Method A: Step (1):

The present compound is dissolved in water, an organic solvent or amixture thereof with a surfactant to prepare a solution of the activeingredient. The resulting solution is sterilely filtered through asterile filter to prepare a sterile solution of the active ingredient.The solvent used herein (water, an organic solvent or a mixture thereof)is preferably an organic solvent or a mixture of an organic solvent andwater, more preferably a mixture of an organic solvent and water.Sterile filters are effective not only for sterilely-filtering, but alsofor removing foreign substances derived from starting materials orforeign substances immixed as a contamination during processing steps.The surfactant includes, for example, polysorbate 80, polysorbate 20,polyoxyethylene hydrogenated castor oil 50, polyoxyethylene hydrogenatedcastor oil 60, poloxamer 188, polyoxyethylene castor oil, benzalkoniumchloride, and sodium lauryl sulfate. In addition, two or more of thesurfactants may be used. Preferably, the surfactant is polysorbate 80.Preferably, the surfactant is contained in about 0.005% (w/v) to about10% (w/v).

The water used herein is purified water or the same or high level ofwater, or water for injection.

The organic solvent includes, for example, alcohol solvents (e.g.methanol, ethanol, etc.), and aprotic solvents (e.g. acetone,dimethylsulfoxide, N,N-dimethylacetamide, etc.). In addition, two ormore of the solvents may be used. Preferably, the organic solvent is1-propanol, methanol, ethanol, 2-propanol, acetone, dimethylsulfoxide,or N,N-dimethylacetamide.

The term “mixture of an organic solvent and water” is defined as a mixedsolvent comprising the above-mentioned alcohol solvent and/or theabove-mentioned aprotic solvent and water (hereinafter, the mixture ofan alcohol solvent and water is optionally referred to as“water-containing alcohol solvent”). The mixture of an organic solventand water includes, for example, a mixture of 1-propanol, methanol,ethanol, 2-propanol, acetone, dimethylsulfoxide, orN,N-dimethylacetamide, and water. The preferable mixture is a mixture ofethanol and water (optionally referred to as “water-containingethanol”).

The organic solvent in the mixture of an organic solvent and water maybe contained in the range of 0% to 100% as long as the organic solventand water can be homogeneously mixed, for example, in 30% to 95%,preferably 40% to 90%, more preferably 50% to 90%, and even morepreferably 60% to 80%. It is especially preferred that the mixture is70% water-containing ethanol. Preferably, the organic solvent iscontained in about 50 to about 90% per the whole volume of thecomposition. The temperature in this step is about 1 to about 90° C.,preferably about 5 to about 60° C.

Step (2):

A water solution comprising a surfactant and a buffer is prepared togive a crystallizing agent. The water solution (crystallizing agent) issterilely filtered through a sterile filter to give a sterilecrystallizing agent, and then the agent is sterilely put into a vesselfor crystallization whose inside is sterilized (hereinafter, optionallyreferred to as a “sealed-type vessel for crystalline” or a “sealedvessel”).

The surfactant includes, for example, polysorbate 80, polysorbate 20,polyoxyethylene hydrogenated castor oil 50, polyoxyethylene hydrogenatedcastor oil 60, poloxamer 188, polyoxyethylene castor oil, benzalkoniumchloride, and sodium lauryl sulfate. Two or more of the surfactants maybe used. The surfactant may be added to the water solution (i.e.crystallizing agent) and/or the sterile solution prepared in step (1)(i.e. sterile solution of the active ingredient). Preferably, thesurfactant is polysorbate 80. Preferably, the surfactant is contained inabout 0.005% (w/v) to about 10% (w/v). The buffer includes, for example,sodium phosphate, sodium dihydrogenphosphate, disodiumhydrogenphosphate, potassium dihydrogenphosphate, dipotassiumhydrogenphosphate, sodium carbonate, triethanolamine, arginine, andmeglumine. Two or more of the buffers may be used. Preferably, thebuffer is sodium dihydrogenphosphate or disodium hydrogenphosphate.Preferably, the buffer is contained in about 0.01% (w/v) to about 8%(w/v). The pH of the water solution is about 5 to about 12, andpreferably about 7 to about 10. The temperature in this step is about 1to about 90° C., and preferably about 5 to about 80° C.

The sealed vessel used in this step includes, for example, a vessel madeof stainless (e.g. SUS304, SUS316, SUS316L) which is equipped with pipearrangement, ferrule, valve, jacket, air filter, stirring system, etc.The stirring system includes, for example, a part of any stirring systemselected from stirring wings (e.g. paddle wings, turbine wings,propeller wings, Pfaudler wings, bar wings, anchor wings), a homomixer,a static mixer and a stirring bar for a magnetic stirrer.

Step (3): Process of Crystallization

The sterile solution prepared in step (1) (sterile solution of theactive ingredient) is sterilely dropped into the vessel forcrystallization (sealed vessel) containing the water solution preparedin step (2) (crystallizing agent). The volume ratio between thesolutions of step (2) and step (1) (i.e. volume ratio between thecrystallizing agent and the solution of the active ingredient) is about1:0.1 to about 1:10, preferably about 1:0.2 to about 1:2.

The present compound is crystallized in this step. The precipitatedcrystal is a cubic crystal wherein the ratio between length and width ofthe cubic crystal is about 1:1, and the ratio between length and heightthereof is about 1:0.8 to about 1:1.2. The mean particle size of thecrystal can be controlled by adjusting the crystallizing temperature,stirring speed, dropping rate, etc., or by adjusting the circulationvelocity when using a bypass pathway mentioned below. The crystallizingtemperature is preferably about 1 to about 90° C., and more preferablyabout 5 to about 60° C. The stirring speed is preferably about 50 rpm toabout 10000 rpm, more preferably 100 rpm to about 5000 rpm, even morepreferably about 100 rpm to about 2000 rpm, and the most preferablyabout 300 rpm to about 1500 rpm. The dropping rate is calculated bydividing the total volume of the sterile solution of step (1) (i.e.sterile solution of the active ingredient) to be added in Step (3) bythe desired time. The dropping rate is, when the total dropped volume ofthe solution of the active ingredient is n L, preferably about n×0.01L/min. to n×1 L/min, more preferably about n×0.01 L/min to about n×0.2L/min, and even more preferably about n×0.05 L/min to about n×0.2 L/min.In more detail, for example, when the total dropped volume of thesolution of the active ingredient is 1 L, the dropping rate is about0.01 L/min to 1 L/min, preferably about 0.01 L/min to about 0.2 L/min,more preferably about 0.05 L/min to about 0.2 L/min; and when the totaldropped volume of the solution of the active ingredient is 100 L, thedropping rate is about 1 L/min to 100 L/min, preferably about 1 L/min toabout 20 L/min, more preferably about 5 L/min to about 20 L/min. And,when the total dropped volume of the solution of the active ingredientis 1000 L, the dropping rate is about 10 L/min to 1000 L/min, preferablyabout 10 L/min to about 200 L/min, more preferably about 50 L/min toabout 200 L/min.

In the step (crystallizing step), the sterile solution (sterile solutionof the active ingredient) prepared in step (1) can be sterilely addeddropwise thereto and simultaneously circulating the solution in thevessel for crystallization via a sealed bypass pathway. Preferably, thebypass pathway has a pump which can pressure a solution in the pathway.Such pump which can pressure a solution includes, for example, a rollerpump (tube pump, hose pump), a reciprocating pump (piston pump, plungerpump, diaphragm pump), and a rotary pump (gear pump, vane pump, screwpump), preferably a roller pump.

The circulation velocity in the bypass pathway is preferably aboutn×0.001 L/min to about n×2 L/min, more preferably about n×0.01 L/min toabout n×1 L/min wherein “n” denotes the total volume of thecrystallizing agent and the solution of the active ingredient. In moredetail, the circulation velocity is about 1 mL/min to about 2000 mL/min,preferably about 10 mL/min to about 1000 mL/min when the total volume ofthe crystallizing agent and the solution of the active ingredient is 1L; about 100 mL/min to about 200 L/min, preferably about 1 L/min toabout 100 L/min when the total volume of the crystallizing agent and thesolution of the active ingredient is 100 L; and about 1 L/min to about2000 L/min, preferably about 10 L/min to about 1000 L/min when the totalvolume of the crystallizing agent and the solution of the activeingredient is 1000 L.

Step (4):

The crystal precipitated in step (3) is collected through a filter inthe sealed vessel. The crystal produced under the condition of in step(3) such as crystallizing temperature and stirring speed has uniformmean particle sizes. The method therefore can prepare a crystal havinguniform mean particle sizes without any milling steps. The sealed vesselused in this step includes, for example, a pressurized vessel made ofstainless (e.g. SUS304, SUS316, SUS316L) which is equipped with pipearrangement, ferrule, valve, crystal collection filter, jacket, airfilter, stirring system, etc. The stirring system includes, for example,a part of any stirring system selected from stirring wings (e.g. paddlewings, turbine wings, propeller wings, Pfaudler wings, bar wings, anchorwings) and a stirring bar for a magnetic stirrer.

Step (5):

A water solution (vehicle solution) comprising a surfactant (about 0.005to about 2% (w/v) per the whole of the final formulation), a buffer(about 0.01 to about 2% (w/v) per the whole of the final formulation),an isotonic agent (about 0.1 to about 10% (w/v) per the whole of thefinal formulation), and water for injection is prepared. The vehiclesolution may further comprise a pH regulator, a soothing agent, apolymer, etc. The prepared solution is sterilely filtered through asterile filter. The resulting vehicle solution is sterilely put into thesealed vessel of step (4) to wash the crystal in the vessel. The washingprocess may be repeated several times. After washing the crystal, thevehicle solution can be sterilely put into the sealed vessel again andmixed with the crystal to provide a suspension formulation which is theformulation of the invention. In addition, the sterile active ingredientprepared in step (4) may be continuously suspended in the vehiclesolution to provide an injectable product (suspension formulation), andfurthermore, the active ingredient obtained from the crystallization maybe washed with water, an organic solvent or a mixture thereof any ofwhich are sterilely filtered through a sterile filter, and then driedfor a while and suspended in the vehicle solution again to prepare asuspension formulation.

The suspension formulation prepared in Step (5) can be formed as aprefilled formulation through the following step (6).

Step (6):

The suspension formulation prepared in Step (5) is sterilely put into acontainer for formulation such as a prefilled syringe, a vial, anampule, a bag or a Blow-Fill-Seal container. The preferred containerincludes a prefilled syringe and a vial.

All of Steps (1) to (6) in the Preparation Method A mentioned above canbe continuously carried out under sterile conditions without taking outthe crystal during the process. According to the present process, it ispossible to easily prepare a sustained release formulation for injectionwithout using the steps of gas sterilization, gamma-ray sterilization,or electron beam sterilization, which is a sterile and dust-freeformulation. In addition, the present process provides asustained-release sterile formulation for injection wherein the activeingredient is uniformly suspended in the solvent.

Preparation Method B:

Preparation Method A can be partially modified as follows. In thefollowing Preparation Method B, the suspension formulation prepared viaStep (1) to Step (5) or Step (6) is not necessarily ensured aboutsterility. In the following Preparation Method B, the embodiments orpreferable embodiments of a surfactant, water, an organic solvent, amixed solvent of an organic solvent and water, a buffer, an isotonicagent, a bypass pathway, a formulation container, etc. used herein canbe the same as Preparation Method A, unless otherwise indicated. Inaddition, pH, temperature, liquid volume ratio, stirring speed, droppingspeed, circulation velocity, etc. used herein are also the same asPreparation Method A, unless otherwise indicated.

Step (1):

The present compound is dissolved in water, an organic solvent or amixture thereof with a surfactant to prepare a solution of the activeingredient.

Step (2):

A water solution comprising a surfactant and a buffer is prepared togive a crystallizing agent. The crystallizing agent is injected into avessel for crystallization.

The vessel for crystallization used herein is equipped with a stirringsystem, whose internal capacity is big enough to contain a solution ofthe active ingredient and the crystallizing agent. The material of thevessel for crystallization includes a stainless vessel (e.g. SUS304,SUS316, SUS316L). The stirring system set in the vessel forcrystallization includes, for example, a part of any stirring systemselected from stirring wings (e.g. paddle wings, turbine wings,propeller wings, Pfaudler wings, bar wings, anchor wings), a homomixer,a static mixer and a stirring bar for a magnetic stirrer.

Step (3): Process of Crystallization

The solution of the active ingredient prepared in step (1) is droppedinto the vessel for crystallization containing the water solutionprepared in step (2) (crystallizing agent). The volume ratio between thecrystallizing agent and the solution of the active ingredient is about1:0.1 to about 1:10, preferably about 1:0.2 to about 1:2.

The present compound is crystallized in this step. The precipitatedcrystal is a cubic crystal wherein the ratio between length and width ofthe cubic crystal is about 1:1, and the ratio between length and heightthereof is about 1:0.8 to about 1:1.2. The mean particle size of thecrystal can be controlled by adjusting the crystallizing temperature,stirring speed, dropping rate, etc., or by adjusting the circulationvelocity when using the bypass pathway mentioned below.

In the step (crystallizing step), the solution of the active ingredientprepared in step (1) can be added dropwise thereto and simultaneouslycirculating the solution in the vessel for crystallization via a bypasspathway. Preferably, the bypass pathway has a pump which can pressure asolution in the pathway. Such pump which can pressure a solutionincludes, for example, a roller pump (tube pump, hose pump), areciprocating pump (piston pump, plunger pump, diaphragm pump), and arotary pump (gear pump, vane pump, screw pump), preferably a rollerpump. Namely, one embodiment of the device which can be used in thecrystallizing process of the present invention includes a deviceequipped with (1) a vessel for crystallization which has a stirring partand can contain a solution of the active ingredient and a crystallizingagent, (2) a bypass pathway connected to the vessel for crystallizationwhich can make a solution in the vessel for crystallization circulated,and (3) a pump set in the bypass pathway which can pressure thesolution. In such device, the bypass pathway can connect, for example,from the vessel for crystallization to the pump, and from the pump tothe vessel for crystallization, with tube, pipe, etc. The tube includes,for example, but not limited to, silicone tube, flexible polyvinylchloride tube, polyurethane tube, fluoro-rubber tube, and olefinic tube,and preferably silicone tube.

The circulation velocity in the bypass pathway is preferably aboutn×0.001 L/min to about n×2 L/min, more preferably about n×0.01 L/min toabout n×1 L/min wherein “n” denotes the total volume of thecrystallizing agent and the solution of the active ingredient. In moredetail, the circulation velocity is about 1 mL/min to about 2000 mL/min,preferably about 10 mL/min to about 1000 mL/min when the total volume ofthe crystallizing agent and the solution of the active ingredient is 1L; about 100 mL/min to about 200 L/min, preferably about 1 L/min toabout 100 L/min when the total volume of the crystallizing agent and thesolution of the active ingredient is 100 L; and about 1 L/min to about2000 L/min, preferably about 10 L/min to about 1000 L/min when the totalvolume of the crystallizing agent and the solution of the activeingredient is 1000 L.

Step (4):

The crystal precipitated in step (3) is collected through a filter. Thecrystal produced under the condition of in step (3) such ascrystallizing temperature, circulation velocity and stirring speed hasalmost uniform mean particle sizes. The method therefore can prepare acrystal having uniform mean particle sizes without any milling steps.

Step (5):

A water solution (vehicle solution) comprising a surfactant, a buffer,an isotonic agent, and water for injection is prepared. The vehiclesolution may further comprise a pH regulator, a soothing agent, awater-soluble polymer, and other polymers, etc. With the resultingvehicle solution, the crystal of step (4) is washed. The washing processmay be repeated several times. After washing the crystal, the vehiclesolution can be put into the vessel again and mixed with the crystal toprovide a suspension formulation which is the formulation of theinvention. In addition, the active ingredient prepared in step (4) maybe continuously suspended in the vehicle solution to provide aninjectable product (suspension formulation), and additionally, theactive ingredient obtained from the crystallization may be washed withwater, an organic solvent or a mixture thereof, and then dried for awhile and suspended in the vehicle solution again to prepare asuspension formulation.

The suspension formulation prepared in Step (5) can be formed as aprefilled formulation through the following step (6).

Step (6):

The suspension formulation prepared in Step (5) is put into a containerfor formulation such as a prefilled syringe, a vial, an ampule, a bag ora Blow-Fill-Seal container. The preferred container includes a prefilledsyringe and a vial.

Step (7):

The prefilled formulation of the suspension (i.e. container filled withthe suspension) prepared in step (6) is sterilized by a process ofsteaming under pressure. The process can make the level of sterileassurance enhanced.

The temperature of the steam sterilization under pressure (high-pressuresteam sterilization) is preferably 100-150° C., more preferably 115-141°C., and even more preferably 115-126° C. The time for the steamsterilization under pressure can be suitably decided, for example, 1-120min, more preferably 5-60 min, even more preferably 10-40 min.

Step (8):

The prefilled formulation of the suspension (i.e. container filled withthe suspension) prepared in step (6) or (7) is ultrasonicated. Theprocess can make the uniformity enhanced to enhance there-suspensiblility when used.

The frequency of the ultrasonication is preferably 10-200 kHz, morepreferably 20-100 kHz, and even more preferably 25-80 kHz. The time forthe ultrasonication can be suitably decided, for example, 0.1-120 min,more preferably 0.1-60 min, even more preferably 0.1-30 min.

It is possible to do any one of the above steps (7) and (8) or both asneeded. When step (7) is carried out, it is preferred to do step (8) touniform the suspension since the active ingredient (free form ofCompound 1) contains macroaggregated particles generated due tosecondary aggregation (see, for example, Example A2, FIGS. 4 and 5mentioned below).

Preparation Method B′

Steps (1)-(6) defined in Preparation Method A are carried out andfurther any one or both of Step (7) and (8) defined in PreparationMethod B are carried out.

According to the above Preparation Method B′ comprising Step (7), thelevel for aseptic assurance of the final product can be ensured sinceSteps (1)-(6) are aseptically carried out, and further the productprepared in Steps (1)-(6) is sterilized by a process of steaming underpressure in Step (7).

In addition, it is also possible to prepare a similar sustained releaseformulation for injection from a physically-milled or spray-driedmaterial of the present compound by means of step (5). Furthermore, theaseptic level of the formulation can be enhanced by carrying out Steps(6), (7) and (8).

Tables 1 and 2 show some typical examples of the sustained releaseformulation for injection. The examples listed in Tables 1 and 2including examples which have combinations of the amount of eachingredient are preferable, but the scope of the present invention is notlimited thereto.

TABLE 1 Ingredients 200 mg/mL 300 mg/mL 400 mg/mL 600 mg/mL Activeingredient 200 mg  300 mg  400 mg  600 mg  (free form of Compound 1)Polysorbate 80 1.7 mg 1.5 mg 1.3 mg 0.9 mg Sodium 1.2 mg 1.0 mg 0.9 mg0.6 mg dihydrogenphosphate(dihydrate) Disodium 2.2 mg 1.9 mg 1.7 mg 1.2mg hydrogenphosphate(anhydride) Sodium chloride 5.8 mg 5.2 mg 4.6 mg 3.2mg Water for injection 1 mL (total volume) pH 7 Osmotic pressure ratio 1Mean particle size 5-25 μm, more preferably 5-20 μm

TABLE 2 Ingredients 50 mg/mL 100 mg/mL 150 mg/mL Active ingredient  50mg 100 mg  150 mg  (free form of Compound 1) Polysorbate 80 2.0 mg 1.9mg 1.8 mg Sodium 1.4 mg 1.3 mg 1.2 mg dihydrogenphosphate(dihydrate)Disodium 2.6 mg 2.4 mg 2.3 mg hydrogenphosphate(anhydride) Sodiumchloride 6.9 mg 6.5 mg 6.2 mg Water for injection 1 mL (total volume) pH7 Osmotic pressure ratio 1 Mean particle size 5-25 μm

EXAMPLES

The present invention is illustrated in more detail by the followingexamples, but it should not be construed to be limited thereto.

Example 1 (Preparation of Formulation No. 1) Step (1):

To an aqueous 70% ethanol solution containing 1% (w/v) polysorbate 80,N-[4-[4-(1,2-benzisothiazol-3-yl)-1-piperazinyl]-(2R,3R)-2,3-tetramethylene-butyl]-(1′R,2′S,3′R,4′S)-2,3-bicyclo[2,2,1]heptanedicarboximidehydrochloride (Compound 1) was dissolved so that the concentration ofCompound 1 should be 2% (w/v), and then the solution was sterilelyfiltered through Millipak™ 60 (model: MPGL06GH2, pore diameter=0.22 μm,MILLIPORE) to prepare 5200 mL of a solution of the active ingredient.

Step (2):

Meanwhile, a water solution of 0.7% (w/v) disodium hydrogenphosphate(anhydride)/1% (w/v) polysorbate 80 was sterilely filtered throughMillipak™ 60 (pore diameter=0.22 μm, MILLIPORE) to prepare 5200 g of acrystallizing agent.

Step (3): Process of Crystallization

To a 1 L glass vessel [model: GLS-80 DURAN™, SCHOTT; with a teflonstirring bar (p 15 mm×70 mm)], 500 g of the crystallizing agent wassterilely injected. Then, maintaining the sealed state of the vessel ata crystallizing temperature of around 25° C., the solution of the activeingredient was added dropwise to the glass vessel at a flow rate of 50mL/min for 10 minutes while stirring the reaction with a magneticstirrer (model: BS-190N, IWAKI) at a stirring speed of 750 rpm to give acrystal slurry of the active ingredient. The crystal in the slurry was acubic-shaped crystal which was a free form of Compound 1.

Step (4):

The crystal slurry was injected to a pressurized filter system (SUS316;model: SF-145, ADVANTEC) under the sealed state to be filtered through asupported PTFE membrane filter pre-attached to the filter system (model:13110014, pore diameter=1 μm, ADVANTEC), then the crystal of the activeingredient was collected on the filter, and the liquid ingredients wereremoved.

Step (5):

To the crystal of the active ingredient which was collected on thefilter, a vehicle solution (about pH 7) comprising sterilely-filtered0.14% (w/v) sodium dihydrogenphosphate (dihydrate)/0.26% (w/v) disodiumhydrogenphosphate (anhydride)/0.70% (w/v) sodium chloride/0.20% (w/v)polysorbate 80 was sterilely injected, and then the mixture wasre-suspended, and filtered. After repeating this procedure severaltimes, the vehicle solution was injected thereto again to prepare thedesired formulation. The injection volume of the vehicle solution wasdecided according to “400 mg/mL” in Table 1 so that each ingredientcould be adapted to the amount calculated based on its density toprepare 400 mg/mL sustained-release formulation for injection defined inTable 1.

Example 2 (Preparation of Formulation No. 2)

According to substantially the same procedure as described in Example 1[Steps (3) to (5)] except that the stirring speed of the magneticstirrer was 675 rpm, using 500 g of the crystallizing agent and 500 mLof the solution of the active ingredient which were prepared in Example1, 400 mg/mL sustained-release sterile formulation defined in Table 1was prepared. The injection volume of the vehicle solution was decidedaccording to “400 mg/mL” in Table 1 so that each ingredient could beadapted to the amount calculated based on its density to prepare 400mg/mL sustained-release formulation for injection defined in Table 1.

Example 3 (Preparation of Formulation No. 3)

According to substantially the same procedure as described in Example 1[Steps (3) to (5)] except that the stirring speed of the magneticstirrer was 400 rpm, using 500 g of the crystallizing agent and 500 mLof the solution of the active ingredient which were prepared in Example1, 400 mg/mL sustained-release sterile formulation defined in Table 1was prepared. The injection volume of the vehicle solution was decidedaccording to “400 mg/mL” in Table 1 so that each ingredient could beadapted to the amount calculated based on its density to prepare 400mg/mL sustained-release formulation for injection defined in Table 1.

Example 4 (Measurement of Particle Size Distribution of Formulations No.1 to No. 3)

Table 3 shows the formulation compositions of Formulations No. 1 to No.3 which were respectively prepared in Examples 1 to 3.

TABLE 3 Formulation composition Active ingredient 400 mg (free form ofCompound 1) Polysorbate 80 1.3 mg Sodium dihydrogenphosphate dihydrate0.9 mg Disodium hydrogenphosphate anhydride 1.7 mg Sodium chloride 4.6mg Water for injection 1 mL (total volume)

(Measurement of Particle Size Distribution)

The particle size distribution was determined as a volume basis with alaser diffraction particle size analyzer (model: SALD-3000, ShimadzuCorporation) by wet method using water as a dispersion medium(refractive index: 2.40-0.20 i). The resulting particle size wasexpressed as a mean particle size (D50) based on its median diameter.

Table 4 shows the particle size distributions (D10, D50, D90) ofFormulations No. 1 to No. 3. These results indicate that it is possibleto control the mean particle size of the crystalline active ingredientby changing the stirring speed of the magnetic stirrer, unless thecrystallizing temperature, and the fluid volumes of the crystallizingagent and the solution of the active ingredient are varied.

TABLE 4 Stirring Particle size distribution (μm) Formulation speed Meanparticle size No. (rpm) D10 D50 D90 1 750 5.4 11.1 19.5 2 675 8.1 15.221.1 3 400 15.0 19.9 25.1

Example 5

To an aqueous 70% ethanol solution containing 1% (w/v) polysorbate 80,Compound 1 was dissolved so that the concentration of Compound 1 shouldbe 2% (w/v), and then the solution was sterilely filtered throughMillipak™ 60 (pore diameter=0.22 μm) to prepare 5200 mL of a solution ofthe active ingredient.

Meanwhile, a water solution of 0.7% (w/v) disodium hydrogenphosphate(anhydride)/1% (w/v) polysorbate 80 was sterilely filtered throughMillipak™ 60 (pore diameter=0.22 μm) to prepare 5000 g of acrystallizing agent.

To a 10 L glass vessel [φ 220 mm×430 mm, caliber φ 95 mm, IWAKI; with ateflon stirring bar for magnetic stirrer (φ 27 mm×108 mm)] whose insideis sterilized, 5000 g of the crystallizing agent was injected. Then, ata crystallizing temperature of around 25° C., the solution of the activeingredient was added dropwise to the glass vessel at a flow rate of 500mL/min for 10 minutes while stirring the reaction with a magneticstirrer (model: BS-190N, IWAKI) at a stirring speed of 650 rpm to give acrystal slurry of the active ingredient. The crystal in the slurry was acubic-shaped crystal which was a free form of Compound 1.

According to substantially the same procedure as described in Example 1[Step (5)], adjusting the concentration of the active ingredient bychanging the injection volume of the vehicle solution, 300 mg/mLsustained-release formulation for injection defined in Table 1 wasprepared.

The particle size distributions were 5.6 μm (D10), 7.6 μm (mean particlesize D50), and 11.7 μm (D90), which were determined by using the methodmeasuring particle size distributions defined in Example 4.

Example 6 (Preparation of Formulation No. 4 and Formulation No. 7)

To an aqueous 70% ethanol solution containing 1% (w/v) polysorbate 80,Compound 1 was dissolved so that the concentration of Compound 1 shouldbe 2% (w/v), and then the solution was sterilely filtered throughMillipak™ 100 (model: MPGL1GCA3, pore diameter=0.22 μm, MILLIPORE) toprepare 5200 mL of a solution of the active ingredient.

Meanwhile, a water solution of 0.7% (w/v) disodium hydrogenphosphate(anhydride)/1% (w/v) polysorbate 80 was sterilely filtered throughMillipak™ 100 (pore diameter=0.22 μm) to prepare 5200 g of acrystallizing agent.

To a 1 L glass bottle [model: GLS-80 DURAN™, SCHOTT; with a teflonstirring bar (φ 15 mm×70 mm)] whose inside is sterilized, 600 g of thecrystallizing agent was sterilely injected. Then, at a crystallizingtemperature of around 1° C. (in ice bath), the solution of the activeingredient was added dropwise to the glass bottle at a flow rate of 60mL/min for 10 minutes while stirring the reaction with a magneticstirrer (model: BS-190N, IWAKI) at a stirring speed of 500 rpm to give acrystal slurry of the active ingredient. The crystal in the slurry was acubic-shaped crystal which was a free form of Compound 1.

According to substantially the same procedure as described in Example 1[Step (5)], adjusting the concentration of the active ingredient bychanging the injection volume of the vehicle solution, 200 mg/mL(Formulation No. 4) and 400 mg/mL (Formulation No. 7) sustained-releaseformulations for injection defined in Table 1 were prepared.

Example 7 (Preparation of Formulations No. 5 and No. 8)

Into a 10 L glass bottle (φ 220 mm×430 mm, caliber φ 95 mm, IWAKI) whoseinside is sterilized, stirring wings (diameter=70 mm, 4 inclined wings,material=SUS304) were sterilely set. Then, under a sterile condition,the crystallizing agent (2500 g) and the solution of the activeingredient (2500 mL) which were prepared in Example 6 were continuouslyadded at a flow rate of 300 mL/min while mixing them with a Y shapedtube (full length=60 mm, internal diameter=6 mm) to the glass bottlewherein the crystallizing temperature was controlled at 25° C. At thetime that about 1 L of the mixture was added thereto (i.e. when thestirring wings started to soak the mixture), the stirring at 200 rpm wasstarted with a stirring machine (model: STIRRER SSR, IWAKI). After allof the crystallizing agent and the solution of the active ingredientwere added while mixing continuously, the reaction was stirred foranother about 15 minutes to give a crystal slurry of the activeingredient. The crystal in the crystal slurry was a cubic-shaped crystalwhich was a free form of Compound 1.

Similarly, according to the same procedure as described in Example 1[Step (5)], 200 mg/mL (Formulation No. 5) and 400 mg/mL (Formulation No.8) sustained-release formulations for injection in Table 1 were preparedby adjusting the volume of the vehicle solution so that theconcentration of the active ingredient could be adapted for the desiredvalue.

Example 8 (Preparation of Formulation No. 6)

To a 2 L glass bottle [model: GLS-80 DURAN™, SCHOTT; with a teflonstirring bar (cp 15 mm×70 mm)] whose inside is sterilized, 1100 g of thecrystallizing agent prepared in Example 6 was sterilely injected. Then,the solution of the active ingredient prepared in Example 7 wassterilely added dropwise at a flow rate of 110 mL/min for 10 minutes tothe glass bottle wherein the crystallizing temperature was controlled at40° C. while stirring with a magnetic stirrer (model: BS-190N, IWAKI) ata stirring speed of 600 rpm to give a crystal slurry of the activeingredient. The crystal in the crystal slurry was a cubic-shaped crystalwhich was a free form of Compound 1.

According to substantially the same procedure as described in Example 1[Step (5)], adjusting the concentration of the active ingredient bychanging the injection volume of the vehicle solution, the 200 mg/mLsustained-release formulation for injection defined in Table 1 wasprepared.

Example 9 (Preparation of Formulation No. 9)

To a 1 L glass bottle [model: GLS-80 DURAN™, SCHOTT; with a teflonstirring bar (φ 15 mm×70 mm)] whose inside is sterilized, 400 g of thecrystallizing agent prepared in Example 6 was sterilely injected. Then,the solution of the active ingredient prepared in Example 6 wassterilely added dropwise at a flow rate of 40 mL/min for 10 minutes tothe glass bottle wherein the crystallizing temperature was controlled at50° C. while stirring with a magnetic stirrer (model: BS-190N, IWAKI) ata stirring speed of 400 rpm to give a crystal slurry of the activeingredient. The crystal in the crystal slurry was a cubic-shaped crystalwhich was a free form of Compound 1.

According to substantially the same procedure as described in Example 1[Step (5)], adjusting the concentration of the active ingredient bychanging the injection volume of the vehicle solution, the 400 mg/mLsustained-release formulation for injection defined in Table 1 wasprepared.

Example 10 (Preparation of Formulation No. 10)

The free form of Compound 1 was milled with a Pin Mill (model AVIS-100,DALTON) at a blade rotational speed of 14000 rpm to give a milled activeingredient. To the milled active ingredient, the sterilely-filteredvehicle solution was sterilely injected and the mixture wasre-suspended, and then the concentration of the active ingredient wasadjusted by changing the injection volume of the vehicle solution togive the 400 mg/mL sustained-release formulation for injection definedin Table 1.

Example 11 (Preparation of Formulation No. 11)

To a 70% ethanol solution containing 1% (w/v) polysorbate 80, Compound 1was dissolved so that the concentration of Compound 1 should be 2%(w/v), and then the solution was sterilely filtered through SFCA BottleTop Filter™ [a bottle-top filter system with cellulose acetate membrane,model: 291-4520, pore diameter=0.22 μm, Nalge Nunc International] toprepare 5000 mL of a solution of the active ingredient.

Meanwhile, a water solution of 0.7% (w/v) disodium hydrogenphosphate(anhydride)/1% (w/v) polysorbate 80 was sterilely filtered through SFCABottle Top Filter™ [pore diameter=0.22 μm] to prepare 5000 g of acrystallizing agent.

To a 300 mL tall beaker [model: TE-32, IWAKI; with a teflon stirring bar(φ 10 mm×35 mm)], 100 mL of the crystallizing agent was injected. Then,the solution of the active ingredient was added dropwise to the beakerat a flow rate of 10 mL/min for 10 minutes while controlling thecrystallizing temperature at 80° C. and stirring with a magnetic stirrer(model: BS-56L-1, IWAKI) at a stirring speed of 1200 rpm to give acrystal slurry of the active ingredient. The crystal in the crystalslurry was a cubic-shaped crystal which was a free form of Compound 1.

According to substantially the same procedure as described in Example 1[Steps (4) to (5)], adjusting the concentration of the active ingredientby changing the injection volume of the vehicle solution, the 400 mg/mLsustained-release sterile formulation defined in Table 1 was prepared,which is also the same as the formulation defined in Table 3.

The particle size distribution was 20.8 μm (D10), 54.3 μm (mean particlesize D50), and 130.3 μm (D90), which were determined by using the methodmeasuring particle size distributions defined in Example 4.

The sterile formulation of the present invention can be prepared throughthe non-sterile process of any one of Examples 1 to 11, and then steamsterilization under pressure and ultrasonication (according to, forexample, the method of Example A1, B3 or B4).

Example A1

To an aqueous 70% ethanol solution containing 1% (w/v) polysorbate 80,Compound 1 was dissolved so that the concentration of Compound 1 shouldbe 2% (w/v), and then the solution was sterilely filtered throughMillipak™ 60 (pore diameter=0.22 μm) to prepare 10500 mL of a solutionof the active ingredient. Meanwhile, a water solution of 0.7% (w/v)disodium hydrogenphosphate (anhydride)/1% (w/v) polysorbate 80 wasfiltered through Millipak™ 60 (pore diameter=0.22 μm) to prepare 10500 gof a crystallizing agent.

To a 10 L glass vessel [φ 220 mm×430 mm, caliber φ 95 mm, IWAKI; with ateflon stirring bar ((27 mm×108 mm)] whose inside is sterilized, 5000 gof the above crystallizing agent was injected. Then, at a crystallizingtemperature of around 25° C., the solution of the active ingredient wasadded dropwise to the glass vessel at a flow rate of 500 mL/min for 10minutes while stirring the reaction with a magnetic stirrer (model:BS-190N, IWAKI) at a stirring speed of 650 rpm to give a crystal slurryof the active ingredient. Using the above 10 L glass vessel, the aboveprocedure was repeated twice to obtain the crystal slurry of the activeingredient whose total amount was 20 L. The crystal in the slurry was acubic-shaped crystal which was a free form of Compound 1.

The crystal slurry was injected to a pressurized filter system (model:SF-145, material=SUS316, ADVANTEC) to be filtered through a supportedPTFE membrane filter pre-attached to the filter system (model: 13110014,pore diameter=1 μm, ADVANTEC), then the crystal of the active ingredientwas collected on the filter, and the liquid ingredients were removed.

To the crystal of the active ingredient which was collected on thefilter, a vehicle solution (about pH 7) comprising sterilely-filtered0.14% (w/v) sodium dihydrogenphosphate (dihydrate)/0.26% (w/v) disodiumhydrogenphosphate (anhydride)/0.70% (w/v) sodium chloride/0.20% (w/v)polysorbate 80 was injected, and then the mixture was re-suspended, andfiltered. After repeating this procedure several times, the vehiclesolution was injected thereto again to prepare the desired formulation.The injection volume of the vehicle solution was decided according toTable 1 so that each ingredient could be adapted to the amountcalculated based on its density to prepare 100, 150 and 200 mg/mLsustained-release formulation for injection defined in Table 1.

Each 2 mL of the sustained release formulations for injection whereinthe concentrations of the active ingredient are 100, 150, and 200 mg/mLwere put into each 4 mL vial (model: V-SC4 mL CS, FUJI GLASS), and thevials were sealed with a rubber stopper and a cap. One mL prefilledsyringe (model: V-OVS System 10.6 rubber formulation 40, VETTER) wasfilled with 1 mL of the sustained release formulation for injectionwherein the concentration of the active ingredient is 150 mg/mL andsealed with a rubber stopper (ST-W 1 mL, VETTER). The vials and theprefilled syringe were sterilized at 121° C. for 20 min with ahigh-pressure steam sterilizer (model: HG-80, HIRAYAMA). Then, the vialsand the prefilled syringe were treated for 3 min with an ultrasonicator(model: UT-605HS, SHARP) at 35 kHz.

The particle size distribution was 5.6 μm (D10), 10.3 μm (mean particlesize D50) and 16.9 μm (D90) in 100 mg/mL formulation laded in vial; 5.6μm (D10), 8.6 μm (mean particle size D50) and 13.1 μm (D90) in 150 mg/mLformulation laded in vial; 5.7 μm (D10), 8.7 μm (mean particle size D50)and 13.9 μm(D90) in 200 mg/mL formulation laded in vial; and 9.9 μm(D10), 22.1 μm (mean particle size D50) and 43.2 μm (D90) in 150 mg/mLformulation laded in prefilled syringe, which were determined by usingthe method measuring particle size distributions defined in Example 4.

Example A2

Using the sustained release formulation for injection (in vial)comprising 150 mg/mL crystalline active ingredient which was juststerilized with a high-pressure steam sterilizer in Example A1, (i)non-ultrasonicated formulation (FIG. 4), and (ii) ultrasonicatedformulation (35 kHz, 3 min) (FIG. 5) were stirred for 30 sec with avortex mixer (model: MT-51, Yamato), and transferred each to 35 mmpolystyrene plate (model: 1000-035, IWAKI). The formulations wereobserved with the naked eye if they contain macroaggregated particles.

The results showed that there were many secondarily-aggregatedmacroaggregated particles in (i) non-ultrasonicated formulation as shownin FIG. 4, while there were no secondarily-aggregated macroaggregatedparticles in (ii) ultrasonicated formulation (35 kHz, 3 min). Thus, likethe ultrasonicated formulation, it became possible to prepare asustained release formulation for injection which has uniform crystal ofan active ingredient.

Example A3 (Powder X-Ray Diffraction)

Using the sustained release formulation for injection (in vial)comprising 150 mg/mL crystalline active ingredient which was prepared inExample A1, the powder X-ray diffraction was measured (i) before thesteam sterilization under pressure, and (ii) after the steamsterilization under pressure (121° C., 20 min) and the ultrasonication(35 kHz, 3 min).

The measurement of powder X-ray diffraction was carried out with anX-ray diffraction measurement device [model: X'pert Pro, CuKα source(λ=1.54056 Å), Kevex solid phase Si(Li) ditector, Spectris], Thecondition of the measurement was Start angle: 5.0°, End angle: 40.00,Step size: 0.017°, Time per step: 5.1 sec, Scan speed: 0.42⁰/sec, X-ray:45 kV, 40 mA, slit (FDS, FASS): ½, 1, mask: 15. The crystal of theactive ingredient was washed with water for injection (OtsukaParmaceutical) and dried in air overnight. The resulting dried crystalpowder was put on the top of a sample holder and the surface thereof wassmoothed with a glass slide.

In the sustained release formulation for injection comprising 150 mg/mLcrystalline active ingredient, the result of X-ray diffraction in thesample before the steam sterilization under pressure is shown in Table 5and FIG. 6, and the result of X-ray diffraction in the sample after thesteam sterilization under pressure (121° C., 20 min) and theultrasonication (35 kHz, 3 min) is shown in Table 6 and FIG. 7.

There was no substantial change in the powder X-ray diffraction betweenthe samples before the steam sterilization under pressure and after thesteam sterilization under pressure and the ultrasonication.

TABLE 5 (before the steam sterilization under pressure) Relatived-spacing Intensity No. 2θ [°] [A] [%] 3 11.2 7.87 38.9 6 14.0 6.33 18.47 15.1 5.86 100.0 8 15.6 5.69 90.3 9 16.3 5.44 73.7 10 16.6 5.34 61.1 1218.1 4.91 86.9 13 19.2 4.63 47.5 14 19.5 4.55 37.7 15 20.0 4.44 98.2 1620.2 4.40 27.3 17 20.7 4.29 23.8 18 20.9 4.25 33.0 19 22.3 3.98 28.7 2223.9 3.72 12.1 23 24.2 3.68 19.7 24 24.7 3.61 17.2 25 24.9 3.58 11.3 2625.4 3.51 25.3 28 26.1 3.42 30.3 29 26.8 3.33 13.7

TABLE 6 [After the steam sterilization under pressure (121° C., 20 min)and the ultrasonication (35 kHz, 3 min)] Relative d-spacing IntensityNo. 2θ [°] [A] [%] 3 11.2 7.91 36.46 6 13.9 6.35 18.06 7 15.1 5.88100.00 8 15.5 5.70 90.50 9 16.3 5.46 73.50 10 16.6 5.35 61.58 12 18.04.92 84.94 13 19.1 4.64 44.32 14 19.5 4.56 38.45 15 20.0 4.45 96.31 1620.2 4.40 24.46 17 20.7 4.30 19.77 18 20.9 4.26 34.27 19 22.3 3.99 29.4621 23.9 3.72 10.77 22 24.1 3.69 21.13 23 24.6 3.61 17.81 24 24.9 3.5811.59 25 25.3 3.52 26.46 27 26.0 3.42 30.81 28 26.8 3.33 11.85

Based on the results of powder X-ray diffraction after the steamsterilization under pressure and the ultrasonication (Table 6 and FIG.7), the specified main and characteristic diffraction peaks are listedbelow. The values of diffraction peaks at diffraction angle 2θ (°) caninclude a little measurement errors according to a measurement device ora measurement condition. For example, the measurement error may be ±0.2,preferably ±0.1.

Main diffraction peaks: 2θ (°)=11.2, 15.1, 15.5, 16.3, 16.6, 18.0, 19.1,19.5, 20.0, 20.9, 22.3, 26.0.

Characteristic diffraction peaks: 2θ (°)=15.1, 15.5, 16.3, 16.6, 18.0,20.0.

Example B1 (without Bypass Pathway)

To an aqueous 70% ethanol solution containing 1% (w/v) polysorbate 80,Compound 1 was dissolved so that the concentration of Compound 1 shouldbe 2% (w/v), and then the solution was sterilely filtered throughMillipak™ 60 (pore diameter=0.22 μm) to prepare 10 L of a solution ofthe active ingredient.

Meanwhile, a water solution of 0.7% (w/v) disodium hydrogenphosphate(anhydride)/1% (w/v) polysorbate 80 was sterilely filtered throughMillipak™ 60 (pore diameter=0.22 μm) to prepare 10 kg of a crystallizingagent.

Into a 1 L glass separable flask (φ 110 mm×180 mm), stirring wings(diameter=70 mm, 4 wings whose inclination is 45°, material=SUS304) wereset. The crystallizing agent (500 mL) was injected into the 1 L glassseparable flask. Then, the solution of the active ingredient was addeddropwise at a flow rate of 20 mL/min to the flask wherein thecrystallizing temperature was controlled at around 25° C. while mixingthem at 400 rpm with a stirring machine (model: FBL1200, ShintoScientific) for 25 min to give a crystal slurry of the activeingredient. The crystal in the crystal slurry was a mixture of acubic-shaped crystal and an amorphous form which was a free form ofCompound 1.

Example B2 (with Bypass Pathway)

Into a 1 L glass separable flask (φ 110 mm×180 mm), stirring wings(diameter=70 mm, 4 inclined wings, material=SUS304) were set, and abypass pathway composed of silicon tube (model: 96410-15, Cole-Parmer),roller pump (model: 7549-32, Cole-Parmer) and pump head (model:77201-62, Cole-Parmer) was set.

The crystallizing agent (500 mL) prepared in Example B1 was injectedinto the 1 L glass separable flask. Then, the solution of the activeingredient prepared in Example B1 was added dropwise at a flow rate of20 mL/min to the flask wherein the crystallizing temperature wascontrolled at around 25° C. while mixing them at 400 rpm with a stirringmachine (model: FBL1200, Shinto Scientific) for 25 min andsimultaneously circulating the solution for 25 min or more at acirculation velocity of 600 mL/min (roller pump, flow rate: 150 mL/min×4pump heads, model: 7523-00, Cole-Parmer) via the above bypass pathway togive a crystal slurry of the active ingredient. The crystal in thecrystal slurry was a cubic-shaped crystal which was a free form ofCompound 1.

Example B3 (with Bypass Pathway)

To an aqueous 70% ethanol solution containing 1% (w/v) polysorbate 80,Compound 1 was dissolved so that the concentration of Compound 1 shouldbe 2% (w/v), and then the solution was sterilely filtered throughMillipak™ 100 (pore diameter=0.22 μm) to prepare 15.5 L of a solution ofthe active ingredient.

Meanwhile, a water solution of 0.7% (w/v) disodium hydrogenphosphate(anhydride)/1% (w/v) polysorbate 80 was filtered through Millipak™ 100(pore diameter=0.22 μm) to prepare 15.5 kg of a crystallizing agent.

Into a 35 L SUS316L vessel (φ 330 mm×550 mm), stirring wings(diameter=120 mm, 3 wings whose inclination is 45°, material=SUS316L,SANKO ASTEC) were set, and a sealed-type bypass pathway composed ofsilicon tube (model: 96410-15, Cole-Parmer), roller pump (model:7549-32, Cole-Parmer) and pump head (model: 77201-62, Cole-Parmer) wasset. The crystallizing agent (15 kg) was injected into the vessel. Then,maintaining the sealed state of the vessel, the solution of the activeingredient was added dropwise at a flow rate of 600 mL/min to the vesselwherein the crystallizing temperature was controlled at around 25° C.while mixing them at 800 rpm (setting pressure: 0.2 MPa) with a stirringmachine with air-motor (model: MRV003A, MAGNEO GIKEN) for 25 min andsimultaneously circulating the solution for 25 min or more at acirculation velocity of 5.4 L/min (roller pump, flow rate: 900 mL/min×6pump heads) via the above bypass pathway to give a crystal slurry of theactive ingredient. The crystal in the crystal slurry was a cubic-shapedcrystal which was a free form of Compound 1.

The substantially same process as Example 1 (Steps 4-5) was done byadjusting the volume of the vehicle solution to adjust the concentrationof the active ingredient, to prepare a sustained release formulation forinjection which contains each ingredient used in the vehicle solutionshown in Table 7 that was calculated based on the density of theformulation and 150 mg/mL crystal of the active ingredient.

Two mL of the sustained release formulations for injection wherein theconcentration of the active ingredient is 150 mg/mL was put into 4 mLvial (model: V-SC4 mL CS, FUJI GLASS), and the vial was sealed with arubber stopper and a cap. The vial was sterilized at 121° C. for 20 minwith a high-pressure steam sterilizer (type: HG-80, HIRAYAMA). Then, thevial was treated for 3 min with an ultrasonicator (model: UT-605HS,SHARP) at 35 kHz.

The particle size distribution was 12.4 μm (D10), 16.7 μm (mean particlesize D50) and 21.9 μm (D90), which were determined by using the methodmeasuring particle size distributions defined in Example 4.

TABLE 7 Composition of formulation Active ingredient 150 mg (free formof Compound 1) Polysorbate 80 1.8 mg Sodium dihydrogenphosphate 1.2 mgdihydrate Disodium hydrogenphosphate 2.3 mg anhydride Sodium chloride6.2 mg Water for injection Total 1 mL

Example B4 (with Bypass Pathway)

To an aqueous 70% ethanol solution containing 1% (w/v) polysorbate 80,Compound 1 was dissolved so that the concentration of Compound 1 shouldbe 2% (w/v), and then the solution was sterilely filtered throughMillipak™ 100 (pore diameter=0.22 μm) to prepare 50 L of a solution ofthe active ingredient.

Meanwhile, a water solution of 0.7% (w/v) disodium hydrogenphosphate(anhydride)/1% (w/v) polysorbate 80 was filtered through Millipak™ 100(pore diameter=0.22 μm) to prepare 50 kg of a crystallizing agent.

Into a 150 L SUS304 vessel (φ 544 mm×630 mm), stirring wings(diameter=120 mm, 3 wings whose inclination is 45°, material=SUS316L,SANKO ASTEC) were set, and a bypass pathway composed of silicon tube(model: 96420-82, Cole-Parmer), roller pump (model: 7549-32,Cole-Parmer) and pump head (model: 77601-10, Cole-Parmer) was set. Thecrystallizing agent (50 kg) was injected into the vessel. Then, thesolution of the active ingredient was added dropwise at a flow rate of 2L/min to the vessel wherein the crystallizing temperature was controlledat around 25° C. while mixing them at 800 rpm (setting pressure: 0.2MPa) with a stirring machine with air-motor (model: MRV003A, MAGNEOGIKEN) for 25 min and simultaneously circulating the solution for 25 minor more at a circulation velocity of 9 L/min (roller pump, flow rate:4.5 L/min×2 pump heads) via the above bypass pathway to give a crystalslurry of the active ingredient. The crystal in the crystal slurry was acubic-shaped crystal which was a free form of Compound 1.

The substantially same process as Example 1 (Steps 4-5) was done byadjusting the volume of the vehicle solution to prepare a sustainedrelease formulation for injection which contains each ingredient used inthe vehicle solution shown in Table 7 that was calculated based on thedensity of the formulation and 150 mg/mL crystal of the activeingredient.

Two mL of the sustained release formulations for injection wherein theconcentration of the active ingredient is 150 mg/mL was put into 4 mLvial (model: V-SC4 mL CS, FUJI GLASS), and the vial was sealed with arubber stopper and a cap. The vial was sterilized at 121° C. for 20 minwith a high-pressure steam sterilizer (type: HG-80, HIRAYAMA). Then, thevial was treated for 3 min with an ultrasonicator (model: UT-605HS,SHARP) at 35 kHz.

The particle size distribution was 14.3 μm (D10), 17.9 μm (mean particlesize D50) and 23.0 μm (D90), which were determined by using the methodmeasuring particle size distributions defined in Example 4.

As mentioned above, the crystallizing process was carried out whilecirculating the solution via the bypass pathway connected to the vesselfor crystallization, and it was possible to reproducibly prepare a cubiccrystal thereof with a pump in the bypass pathway such as a roller pump(tube pump, hose pump), a reciprocating pump (piston pump, plunger pump,diaphragm pump), and a rotary pump (gear pump, vane pump, screw pump)even when a large-scale preparation.

Example C1 (Effect of Ethanol in Solution of the Active Ingredient) (70%Ethanol-Containing Solution of the Active Ingredient)

To an aqueous 70% ethanol solution containing 1% (w/v) polysorbate 80 asa water-containing alcohol solvent, Compound 1 was dissolved so that theconcentration of Compound 1 should be 2% (w/v), and then the mixture waswell stirred to be dissolved as far as possible. Then, the solution wassterilely filtered through SFCA Bottle Top Filter™ (pore diameter=0.22μm) to prepare 1000 mL of a 70% ethanol solution of the activeingredient.

Meanwhile, a water solution of 0.7% (w/v) disodium hydrogenphosphate(anhydride)/1% (w/v) polysorbate 80 was sterilely filtered through SFCABottle Top Filter™ (pore diameter=0.22 μm) to prepare 10 kg of acrystallizing agent.

Into a 1 L glass separable flask (p 110 mm×180 mm), stirring wings(diameter=70 mm, 4 inclined wings, material=SUS304) were set, and abypass pathway composed of silicon tube (model: 96410-15, Cole-Parmer),roller pump (model: 7549-32, Cole-Parmer) and pump head (model:77201-62, Cole-Parmer) was set. The crystallizing agent (500 mL) wasinjected into the 1 L glass separable flask. Then, the solution of theactive ingredient was added dropwise at a flow rate of 20 mL/min to theflask wherein the crystallizing temperature was controlled at around 25°C. while mixing them at 400 rpm with a stirring machine (model: FBL1200,Shinto Scientific) for 25 min and simultaneously circulating thesolution for 25 min or more at a circulation velocity of 600 mL/min(roller pump, flow rate: 150 mL/min×4 pump heads, model: 7523-00,Cole-Parmer) via the above bypass pathway to give a crystal slurry ofthe active ingredient. The crystal in the crystal slurry was acubic-shaped crystal which was a free form of Compound 1.

The crystal slurry was injected to a pressurized filter system (SUS316;model SF-145, ADVANTEC) to be filtered through a supported PTFE membranefilter pre-attached to the filter system (model: 13110014, porediameter=1 μm, ADVANTEC), then the crystal of the active ingredient wascollected on the filter, and the liquid ingredients were removed.

Water for injection (Otsuka Pharmaceutical) was injected on the crystalof the active ingredient collected on the filter, the crystal wasre-suspended, and then the resulting crystal was collected. Theprocedure was repeated several times to wash the crystal of the activeingredient. The crystal of the active ingredient collected on thesupported PTFE membrane filter was dried at 60° C. for 4 hours, theweight of the crystal was weighed (10.51 g, yield: 97.3% from 10.8 g ofthe starting material).

The particle size distribution was 5.5 μm (D10), 10.5 μm (mean particlesize D50) and 18.9 μm (D90), which were determined by using the methodmeasuring particle size distributions defined in Example 4.

Examples C2 to C11

The substantially same process as Example C1 was done by changing theconcentration of ethanol used as a water-containing alcohol solvent from0% to 100% to prepare a cubic-shaped crystal of the active ingredient.The weight of the resulting the crystal of the active ingredient (driedweight), the yield per 10.8 g of the starting material, and the particlesize distribution of the resulting formulations which was measuredaccording to the method of Example 4 are shown in Table 8.

TABLE 8 Concentration of ethanol in the Weight of the Particle sizesolution of the precipitated Yield per the Particle size distributionParticle size active crystal starting distribution (D50; meandistribution Example No. ingredient (%) (yield)/g material/% (D10)/μmparticle size)/μm (D90)/μm C2 0 1.48 13.7 0.8 3.5 6.6 C3 10 1.62 15.02.3 5.6 10.9 C4 20 2.16 20.0 3.9 8.8 17.9 C5 30 3.42 31.7 3.3 7.4 12.1C6 40 6.27 58.1 5.6 8.4 13.0 C7 50 9.63 89.2 6.0 8.7 12.7 C8 60 10.3896.1 5.6 9.4 17.5 C1 70 10.51 97.3 5.5 10.5 18.9 C9 80 10.12 93.7 6.613.0 21.1 C10 90 6.03 55.8 5.4 10.9 19.4 C11 100 1.14 10.6 5.0 12.5 24.1

Based on the above-listed results of each particle size of the preparedcubic crystal and each yield, the content of an organic solvent in themixture of an organic solvent and water to prepare the solution of theactive ingredient was preferably 40 to 90%, more preferably 60 to 80%.In particular, aqueous 70% ethanol solution was the most preferred.

Example D1

To an aqueous 70% ethanol solution containing 1% (w/v) polysorbate 80,Compound 1 was dissolved so that the concentration of Compound 1 shouldbe 2% (w/v), and then the solution was sterilely filtered throughMillipak™ 60 (pore diameter=0.22 μm) to prepare 5200 mL of a solution ofthe active ingredient.

Meanwhile, a water solution of 0.7% (w/v) disodium hydrogenphosphate(anhydride)/1% (w/v) polysorbate 80 was sterilely filtered throughMillipak™ 60 (pore diameter=0.22 μm) to prepare 5200 g of acrystallizing agent.

To a 10 L glass vessel [φ 220 mm×430 mm, caliber φ 95 mm, IWAKI; with ateflon stirring bar for magnetic stirrer (φ 27 mm×108 mm)] whose insideis sterilized, 5000 g of the crystallizing agent was injected. Then, ata crystallizing temperature of around 25° C., the solution of the activeingredient was added dropwise to the glass vessel at a flow rate of 500mL/min for 10 minutes while stirring the reaction with a magneticstirrer (model BS-190N, IWAKI) at a stirring speed of 600 rpm to give acrystal slurry of the active ingredient. The crystal in the slurry was acubic-shaped crystal which was a free form of Compound 1.

The crystal slurry was injected to a SUS316 pressurized filter system(model SF-145, ADVANTEC) to be filtered through a supported PTFEmembrane filter pre-attached to the filter system (model: 13110014, porediameter=1 μm, ADVANTEC), then the crystal of the active ingredient wascollected on the filter, and the liquid ingredients were removed.

To the crystal of the active ingredient which was collected on thefilter, a sterilely-filtered vehicle solution (about pH 7) comprising0.14% (w/v) sodium dihydrogenphosphate (dihydrate)/0.26% (w/v) disodiumhydrogenphosphate (anhydride)/0.70% (w/v) sodium chloride/0.005% (w/v)polysorbate 80 was injected, and then the mixture was re-suspended, andfiltered. After repeating this procedure several times, the vehiclesolution was injected thereto again to prepare the desired formulation.The injection volume of the vehicle solution was decided according to“300 mg/mL” in Table 1 so that each ingredient could be adapted to theamount calculated based on its density to prepare 300 mg/mLsustained-release formulation for injection defined in Table 1. Theparticle size distribution was 9.0 μm (D10), 18.3 μm (mean particle sizeD50) and 30.6 μm (D90), which were determined by using the methodmeasuring particle size distributions defined in Example 4.

In addition, the volume of the vehicle solution can be adjusted toprepare a formulation of the active ingredient in any concentrationbesides 300 mg/mL (50, 100, 150, 200, 250, 400, 500, 600 mg/mL,etc.)[Example D1a].

Examples D2−D9

The substantially same process as Example D1 was done by using thecrystal of the active ingredient prepared in Example D1 and changing theconcentration of polysorbate 80 used in the vehicle solution from 0.01%(w/v) to 0.10% (w/v) to prepare a sustained release formulation forinjection which contains 300 mg/mL cubic-shaped crystal of the activeingredient [Examples D2-D5]. The particle size distribution of theresulting formulations which was measured according to the method ofExample 4 is shown in Table 9.

Likewise, the substantially same process as Example D1 was done bychanging the concentration of polysorbate 80 used in the vehiclesolution from 0.5% (w/v) to 2.0% (w/v) to prepare a sustained releaseformulation for injection which contains a cubic-shaped crystal of theactive ingredient [Examples D6-D9; Table 10].

In addition, the volume of the vehicle solution can be adjusted toprepare a formulation of the active ingredient in any concentrationbesides 300 mg/mL (50, 100, 150, 200, 250, 400, 500, 600 mg/mL, etc.)(Examples D2a-D9a; Table 11).

TABLE 9 Concentration of polysorbate 80 Concentration of Particle sizein the vehicle the active Particle size distribution Particle sizesolution ingredient distribution (D50; mean distribution Example No. (%(w/v)) (mg/mL) (D10)/μm particle size)/μm (D90)/μm D1 0.005 300 9.0 18.330.6 D2 0.01 300 8.4 17.6 29.5 D3 0.02 300 7.3 13.8 23.4 D4 0.05 300 7.513.5 20.1 D5 0.10 300 7.2 12.1 20.4

TABLE 10 Concentration of polysorbate 80 in the Concentration of theExample vehicle solution active ingredient No. (% (w/v)) (mg/mL) D6 0.5300 D7 1.0 300 D8 1.5 300 D9 2.0 300

TABLE 11 Concentration of polysorbate 80 in the Concentration Examplevehicle solution of the active No. (% (w/v)) ingredient (mg/mL) D1a0.005 50, 100, 150, 200, 250, 400, 500, 600, etc. D2a 0.01 50, 100, 150,200, 250, 400, 500, 600, etc. D3a 0.02 50, 100, 150, 200, 250, 400, 500,600, etc. D4a 0.05 50, 100, 150, 200, 250, 400, 500, 600, etc. D5a 0.1050, 100, 150, 200, 250, 400, 500, 600, etc. D6a 0.5 50, 100, 150, 200,250, 400, 500, 600, etc. D7a 1.0 50, 100, 150, 200, 250, 400, 500, 600,etc. D8a 1.5 50, 100, 150, 200, 250, 400, 500, 600, etc. D9a 2.0 50,100, 150, 200, 250, 400, 500, 600, etc.

Example E1

Step (1): To an aqueous 70% ethanol solution containing 1% (w/v)polysorbate 20, Compound 1 was dissolved so that the concentration ofCompound 1 should be 2% (w/v), and then the solution was sterilelyfiltered through SFCA Bottle Top Filter™ (pore diameter=0.22 μm) toprepare 120 mL of a solution of the active ingredient.Step (2): Meanwhile, a water solution of 0.7% (w/v) disodiumhydrogenphosphate (anhydride)/1% (w/v) polysorbate 20 was sterilelyfiltered through SFCA Bottle Top Filter™ (pore diameter=0.22 μm) toprepare 120 mL of a crystallizing agent.Step (3): To a 100 mL vial container with a scale attachment [model:WT-110, NICHIDEN-RIKA GLASS; with a teflon stirring bar (φ 8 mm×35 mm)],50 mL of the crystallizing agent was injected. Then, at a crystallizingtemperature of around 25° C., the solution of the active ingredient wasadded dropwise into the glass bottle at a flow rate of 5 mL/min for 10minutes while stirring the reaction with a magnetic stirrer (modelBS-56L-1, IWAKI) at a stirring speed of 1200 rpm to give a crystalslurry of the active ingredient. The crystal in the slurry was acubic-shaped crystal which was a free form of Compound 1.Step (4): The crystal slurry was injected to a reduced-pressure-typeglass filter system (index No. XX1504700, MILLIPORE) to be filteredthrough Omnipore™ membrane filter pre-attached to the filter system(model: JAWP, pore diameter=1 μm, MILLIPORE), then the crystal of theactive ingredient was collected on the filter, and the liquidingredients were removed.Step (5): To the crystal of the active ingredient which was collected onthe filter, a sterilely-filtered vehicle solution (about pH 7)comprising 0.14% (w/v) sodium dihydrogenphosphate (dihydrate)/0.26%(w/v) disodium hydrogenphosphate (anhydride)/0.70% (w/v) sodiumchloride/0.20% (w/v) polysorbate 80 was injected, and then the mixturewas re-suspended, and filtered. After repeating this procedure severaltimes, the vehicle solution was injected thereto again to prepare thedesired formulation. A sustained-release formulation for injectioncontaining 200 mg/mL the active ingredient was prepared by adjusting thevolume of the vehicle solution. The particle size distribution was 7.5μm (D10), 9.8 μm (mean particle size D50) and 13.1 μm (D90), which weredetermined by using the method measuring particle size distributionsdefined in Example 4.

In addition, the volume of the vehicle solution can be adjusted toprepare a formulation of the active ingredient in any concentrationbesides 200 mg/mL (50, 100, 150, 250, 300, 400, 500, 600 mg/mL, etc.)[Example E1a].

Furthermore, the polysorbate 80 contained in the vehicle solution can bereplaced with polysorbate 20 and the volume of the vehicle solution canbe adjusted to prepare a formulation of the active ingredient in anyconcentration (50, 100, 150, 200, 250, 300, 400, 500, 600 mg/mL, etc.)[Example E1b].

Examples E2-E8

The surfactant (polysorbate 20) used in the solution of the activeingredient (Step (1)) and the crystallizing agent (Step (2)) in ExampleE1 were replaced with any other surfactant in the same concentrationsuch as polyoxyethylene hydrogenated castor oil 50 [Example E2],polyoxyethylene hydrogenated castor oil 60 [Example E3], poloxamer 188[Example E4], polyoxyethylene castor oil [Example E5], benzalkoniumchloride [Example E6], sodium lauryl sulfate [Example E7], andadditive-free (0%) [Example E8], and then the substantially same processas Example E1 was done to prepare a sustained release formulation forinjection which contains 200 mg/mL cubic-shaped crystal of the activeingredient. The surfactant used in each example and the particle sizedistribution of the resulting formulations which was measured accordingto the method of Example 4 are shown in Table 12.

In addition, the volume of the vehicle solution can be adjusted toprepare a formulation of the active ingredient in any concentrationbesides 200 mg/mL (50, 100, 150, 250, 300, 400, 500, 600 mg/mL,etc.)(Table 13).

In addition, the polysorbate 80 used in the vehicle solution (Step (5))in Example E1 was replaced with any other surfactant in the sameconcentration such as polyoxyethylene hydrogenated castor oil 50[Example E2b], polyoxyethylene hydrogenated castor oil 60 [Example E3b],poloxamer 188 [Example E4b], polyoxyethylene castor oil [Example E5b],benzalkonium chloride [Example E6b], and sodium lauryl sulfate [ExampleE7b], and then the substantially same process as Step (5) in Example E1was done adjusting the volume of the vehicle solution to prepare aformulation which contains a cubic-shaped crystal of the activeingredient in any concentration (50, 100, 150, 200, 250, 300, 400, 500,600 mg/mL, etc.) (Table 14).

TABLE 12 Surfactant in the solution of the active Surfactant in theingredient and vehicle solution in Concentration of the Particle sizecrystallizing agent Step (5) active ingredient Particle sizedistribution (D50; mean Particle size Example No. (each 1%(w/v) (each0.2% (w/v) (mg/mL) distribution (D10)/μm particle size)/μm distribution(D90)/μm E1 polysorbate 20 polysorbate 200 7.5 9.8 13.1 80 E2polyoxyethylene polysorbate 200 11.9 15.1 19.5 hydrogenated 80 castoroil 50 E3 polyoxyethylene polysorbate 200 9.8 12.3 15.7 hydrogenated 80castor oil 60 E4 poloxamer 188 polysorbate 200 8.1 15.1 24.8 80 E5polyoxyethylene polysorbate 200 7.9 13.7 19.2 castor oil 80 E6benzalkonium polysorbate 200 9.2 12.9 19.0 chloride 80 E7 sodium laurylpolysorbate 200 10.0 15.3 21.0 sulfate 80 E8 additive-free polysorbate200 53.1 180.6 284.4 (0%) 80

TABLE 13 Surfactant in the solution of Surfactant in the active thevehicle ingredient and solution in Concentration crystallizing Step (5)of the active Example agent (each 1% (each 0.2% ingredient No. (w/v)(w/v) (mg/mL) E1a polysorbate 20 polysorbate 80 50, 100, 150, 250, 300,400, 500, 600, etc. E2a polyoxyethylene polysorbate 80 50, 100, 150,hydrogenated 250, 300, 400, castor oil 50 500, 600, etc. E3apolyoxyethylene polysorbate 80 50, 100, 150, hydrogenated 250, 300, 400,castor oil 60 500, 600, etc. E4a poloxamer 188 polysorbate 80 50, 100,150, 250, 300, 400, 500, 600, etc. E5a polyoxyethylene polysorbate 8050, 100, 150, castor oil 250, 300, 400, 500, 600, etc. E6a benzalkoniumpolysorbate 80 50, 100, 150, chloride 250, 300, 400, 500, 600, etc. E7asodium lauryl polysorbate 80 50, 100, 150, sulfate 250, 300, 400, 500,600, etc.

TABLE 14 Surfactant in the solution of the active Surfactant iningredient and the vehicle crystallizing solution in Step Concentrationof Example agent (each 1% (5) the active No. (w/v) (each 0.2% (w/v)ingredient (mg/mL) E1b polysorbate 20 polysorbate 20 50, 100, 150, 200,250, 300, 400, 500, 600, etc. E2b polyoxyethylene polyoxyethylene 50,100, 150, 200, hydrogenated hydrogenated 250, 300, 400, castor oil 50castor oil 50 500, 600, etc. E3b polyoxyethylene polyoxyethylene 50,100, 150, 200, hydrogenated hydrogenated 250, 300, 400, castor oil 60castor oil 60 500, 600, etc. E4b poloxamer 188 poloxamer 188 50, 100,150, 200, 250, 300, 400, 500, 600, etc. E5b polyoxyethylenepolyoxyethylene 50, 100, 150, 200, castor oil castor oil 250, 300, 400,500, 600, etc. E6b benzalkonium benzalkonium 50, 100, 150, 200, chloridechloride 250, 300, 400, 500, 600, etc. E7b sodium lauryl sodium lauryl50, 100, 150, 200, sulfate sulfate 250, 300, 400, 500, 600, etc.

Example F1

To an aqueous 70% ethanol solution containing 1% (w/v) polysorbate 80,Compound 1 was dissolved so that the concentration of Compound 1 shouldbe 2% (w/v), and then the solution was sterilely filtered through SFCABottle Top Filter™ (pore diameter=0.22 μm) to prepare 500 mL of asolution of the active ingredient.

Meanwhile, a water solution of 50 mmol/L dipotassiumhydrogenphosphate/1% (w/v) polysorbate 80 was sterilely filtered throughSFCA Bottle Top Filter™ (pore diameter=0.22 μm) to prepare 500 g of acrystallizing agent.

To a 100 mL vial container with a scale attachment [model: WT-110,NICHIDEN-RIKA GLASS; with a teflon stirring bar (φ 8 mm×35 mm)], 50 mLof the crystallizing agent was injected. Then, at a crystallizingtemperature of around 25° C., the solution of the active ingredient wasadded dropwise into the glass bottle at a flow rate of 5 mL/min for 10minutes while stirring the reaction with a magnetic stirrer (modelBS-56L-1, IWAKI) at a stirring speed of 1200 rpm to give a crystalslurry of the active ingredient. The crystal in the slurry was acubic-shaped crystal which was a free form of Compound 1.

The crystal slurry was injected to a reduced-pressure-type glass filtersystem (index No. XX1504700, MILLIPORE) to be filtered through Omnipore™membrane filter pre-attached to the filter system (model: JAWP, porediameter=1 μm, MILLIPORE), then the crystal of the active ingredient wascollected on the filter, and the liquid ingredients were removed.

To the crystal of the active ingredient which was collected on thefilter, a sterilely-filtered vehicle solution (about pH 7) comprising0.14% (w/v) sodium dihydrogenphosphate (dihydrate)/0.26% (w/v) disodiumhydrogenphosphate (anhydride)/0.70% (w/v) sodium chloride/0.2% (w/v)polysorbate 80 was injected, and then the mixture was re-suspended, andfiltered. After repeating this procedure several times, the vehiclesolution was injected thereto again to prepare the desired formulation.The injection volume of the vehicle solution was decided according to“200 mg/mL” in Table 1 so that each ingredient could be adapted to theamount calculated based on its density to prepare 200 mg/mLsustained-release formulation for injection defined in Table 1. Theparticle size distribution was 6.3 μm (D10), 10.2 μm (mean particle sizeD50), 16.7 μm (D90), which were determined by using the method measuringparticle size distributions defined in Example 4.

In addition, the volume of the vehicle solution can be adjusted toprepare a formulation of the active ingredient in any concentrationbesides 200 mg/mL (50, 100, 150, 250, 300, 400, 500, 600 mg/mL, etc.)[Example F1a].

Furthermore, the 0.14% (w/v) sodium dihydrogenphosphate(dihydrate)/0.26% (w/v) disodium hydrogenphosphate (anhydride) containedin the vehicle solution can be replaced with potassiumdihydrogenphosphate/dipotassium hydrogen phosphate in order to adjust pHto around 7, and the volume of the vehicle solution can be adjusted toprepare a formulation of the active ingredient in any concentration (50,100, 150, 200, 250, 300, 400, 500, 600 mg/mL, etc.) [Example F1b].

Examples F2-F8

The 50 mmol/L dipotassium hydrogen phosphate (buffer) used in thecrystallizing agent in Example F1 was replaced with any other buffer inthe same mole concentration (mol/L) or a pH regulator such as arginine[Example F2], trometamol [Example F3], sodium carbonate [Example F4],triethanolamine [Example F5], meglumine [Example F6], sodium hydroxide[Example F7], and disodium hydrogenphosphate [Example F8]) and then thesubstantially same process as Example F1 was done to prepare a sustainedrelease formulation for injection which contains 200 mg/mL cubic-shapedcrystal of the active ingredient. The buffer used in each example andthe particle size distribution of the resulting formulations which wasmeasured according to the method of Example 4 are shown in Table 15.

In addition, the volume of the vehicle solution can be adjusted toprepare a formulation of the active ingredient in any concentrationbesides 200 mg/mL (50, 100, 150, 250, 300, 400, 500, 600 mg/mL,etc.)(Table 16).

In addition, the buffer [0.14% (w/v) sodium dihydrogenphosphate(dihydrate)/0.26% (w/v) disodium hydrogenphosphate (anhydride)] used inthe vehicle solution in Example F1 was replaced with any other buffersuch as L-arginine hydrochloride/L-arginine (pH: about 7) [Example F2b],trometamol (pH: about 7) [Example F3b], sodium hydrogen carbonate/sodiumcarbonate (pH: about 7) [Example F4b], triethanolamine (pH: about 7)[Example F5b], and meglumine (pH: about 7) [Example F6b]), and then thesubstantially same process as Example F1 was done adjusting the volumeof the vehicle solution to prepare a formulation which contains acubic-shaped crystal of the active ingredient in any concentration (50,100, 150, 200, 250, 300, 400, 500, 600 mg/mL, etc.) (Table 17).

TABLE 15 Buffer in the Concentration of Particle size crystallizingBuffer in the the active Particle size distribution Particle size agent(each 50 vehicle ingredient distribution (D50; mean distribution ExampleNo. mmol/L) solution (mg/mL) (D10)/μm article size)/μm (D90)/μm F1dipotassium hydrogen 0.14% (w/v) sodium 200 6.3 10.2 16.7 phosphatedihydrogenphosphate (dihydrate)/0.26% (w/v) disodium hydrogenphosphate(anhydride) F2 arginine same as above 200 8.1 10.2 13.4 F3 trometamolsame as above 200 7.1 10.5 19.4 F4 sodium same as above 200 7.2 9.6 14.8carbonate F5 triethanol- same as above 200 8.0 10.5 14.8 amine F6meglumine same as above 200 6.7 9.0 15.8 F7 sodium same as above 20019.2 25.6 33.4 hydroxide F8 disodium same as above 200 6.3 10.2 16.7hydrogen- phosphate

TABLE 16 Buffer in the Concentration crystallizing of the active Exampleagent Buffer in the ingredient No. (each 50 mmol/L) vehicle solution(mg/mL) F1a dipotassium 0.14% (w/v)sodium 50, 100, 150, hydrogendihydrogenphosphate 250, 300, 400, phosphate (dihydrate)/0.26% 500, 600,etc. (w/v)disodium hydrogenphosphate (anhydride) F2a arginine same asabove 50, 100, 150, 250, 300, 400, 500, 600, etc. F3a trometamol same asabove 50, 100, 150, 250, 300, 400, 500, 600, etc. F4a sodium carbonatesame as above 50, 100, 150, 250, 300, 400, 500, 600, etc. F5atriethanolamine same as above 50, 100, 150, 250, 300, 400, 500, 600,etc. F6a meglumine same as above 50, 100, 150, 250, 300, 400, 500, 600,etc. F7a sodium hydroxide same as above 50, 100, 150, 250, 300, 400,500, 600, etc. F8a disodium same as above 50, 100, 150,hydrogenphosphate 250, 300, 400, 500, 600, etc.

TABLE 17 Buffer in the crystallizing Concentration of Example agentBuffer in the the active No. (each 50 mmol/L) vehicle solutioningredient (mg/mL) F1b dipotassium potassium 50, 100, 150, 200, hydrogendihydrogen- 250, 300, 400, phosphate phosphate/ 500, 600, etc.dipotassium hydrogen phosphate (pH about 7) F2b arginine L-arginine 50,100, 150, 200, hydrochloride/L- 250, 300, 400, arginine 500, 600, etc.(pH about 7) F3b trometamol trometamol 50, 100, 150, 200, (pH about 7)250, 300, 400, 500, 600, etc. F4b sodium carbonate sodium hydrogen 50,100, 150, 200, carbonate/sodium 250, 300, 400, carbonate 500, 600, etc.(pH about 7) F5b triethanolamine triethanolamine 50, 100, 150, 200, (pHabout 7) 250, 300, 400, 500, 600, etc. F6b meglumine meglumine 50, 100,150, 200, (pH about 7) 250, 300, 400, 500, 600, etc.

Examples F9 to F16

The 0.70% (w/v) sodium chloride (isotonic agent) used in the vehiclesolution in Example F1 can be replaced with any other isotonic agentsuch as D-mannitol [about 5% (w/v); Example F9], D-fructose [about 5%(w/v); Example F10], D-xylitol [about 5% (w/v); Example F11], D-sorbitol[about 5% (w/v); Example F12], D-lactose [about 10% (w/v); Example F13],D-glucose [about 5% (w/v); Example F14], D-trehalose [about 10% (w/v);Example F15], and D-sucrose [about 10% (w/v); Example F16] and D-maltose[about 5% (w/v); Example F17]) to prepare a sustained releaseformulation for injection of the present invention. The volume of thevehicle solution can be adjusted to prepare a formulation of the activeingredient in any concentration (50, 100, 150, 200, 250, 300, 400, 500,600 mg/mL, etc.) (Table 18).

TABLE 18 Concentration of Isotonic agent isotonic agent (perConcentration of Example in the vehicle the vehicle the active No.solution solution) % (w/v) ingredient (mg/mL) F1 sodium chloride 0.7050, 100, 150, 200, 250, 300, 400, 500, 600, etc. F9 D-mannitol about 550, 100, 150, 200, 250, 300, 400, 500, 600, etc. F10 D-fructose about 550, 100, 150, 200, 250, 300, 400, 500, 600, etc. F11 D-xylitol about 550, 100, 150, 200, 250, 300, 400, 500, 600, etc. F12 D-sorbitol about 550, 100, 150, 200, 250, 300, 400, 500, 600, etc. F13 D-lactose about 1050, 100, 150, 200, 250, 300, 400, 500, 600, etc. F14 D-glucose about 550, 100, 150, 200, 250, 300, 400, 500, 600, etc. F15 D-trehalose about10 50, 100, 150, 200, 250, 300, 400, 500, 600, etc. F16 D-sucrose about10 50, 100, 150, 200, 250, 300, 400, 500, 600, etc. F17 D-maltose about10 50, 100, 150, 200, 250, 300, 400, 500, 600, etc.

Example F18

Lidocaine hydrochloride [about 0.5% (w/v)−2% (w/v)] can be additionallycontained in the vehicle solution prepared in Example F8 and the volumeof the vehicle solution can be adjusted to prepare a formulation oflidocaine hydrochloride in any concentration (50, 100, 150, 200, 250,300, 400, 500, 600 mg/mL, etc.).

Example F19

Lidocaine hydrochloride described in Example F17 can be replaced withprocaine hydrochloride [about 0.5% (w/v)-2% (w/v)] and the volume of thevehicle solution can be adjusted to prepare a formulation of procainehydrochloride in any concentration (50, 100, 150, 200, 250, 300, 400,500, 600 mg/mL, etc.).

Example F20

Lidocaine hydrochloride described in Example F17 can be replaced withmeprylcaine hydrochloride [about 1% (w/v)-3% (w/v)] and the volume ofthe vehicle solution can be adjusted to prepare a formulation ofmeprylcaine hydrochloride in any concentration (50, 100, 150, 200, 250,300, 400, 500, 600 mg/mL, etc.).

Example G1 (Effect of Solvent in Solution of the Active Ingredient)

To an aqueous 70% methanol solution containing 1% (w/v) polysorbate 80,Compound 1 was dissolved so that the concentration of Compound 1 shouldbe 2% (w/v), and then the solution was sterilely filtered through SFCABottle Top Filter™ (pore diameter=0.22 μm) to prepare 120 mL of asolution of the active ingredient.

Meanwhile, a water solution of 0.7% (w/v) disodium hydrogenphosphate(anhydride)/1% (w/v) polysorbate 20 was sterilely filtered through SFCABottle Top Filter™ (pore diameter=0.22 μm) to prepare 120 mL of acrystallizing agent.

To a 100 mL vial container with a scale attachment [model: WT-110,NICHIDEN-RIKA GLASS; with a teflon stirring bar (φ 8 mm×35 mm)], 50 mLof the crystallizing agent was injected. Then, at a crystallizingtemperature of around 25° C., the solution of the active ingredient wasadded dropwise into the glass bottle at a flow rate of 5 mL/min for 10minutes while stirring the reaction with a magnetic stirrer (modelBS-56L-1, IWAKI) at a stirring speed of 1200 rpm to give a crystalslurry of the active ingredient. The crystal in the slurry was acubic-shaped crystal which was a free form of Compound 1.

The crystal slurry was injected to a reduced-pressure-type glass filtersystem (index No. XX1504700, MILLIPORE) to be filtered through Omnipore™membrane filter pre-attached to the filter system (model: JAWP, porediameter=1 μm, MILLIPORE), then the crystal of the active ingredient wascollected on the filter, and the liquid ingredients were removed.

To the crystal of the active ingredient which was collected on thefilter, a sterilely-filtered vehicle solution (about pH 7) comprising0.14% (w/v) sodium dihydrogenphosphate (dihydrate)/0.26% (w/v) disodiumhydrogenphosphate (anhydride)/5.0% (w/v) D-mannitol/0.2% (w/v)polysorbate 80 was injected, and then the mixture was re-suspended, andfiltered. After repeating this procedure several times, the vehiclesolution was injected thereto again to prepare the desired formulation.The injection volume of the vehicle solution was decided according to“200 mg/mL” in Table 1 so that each ingredient could be adapted to theamount calculated based on its density to prepare 200 mg/mLsustained-release formulation for injection defined in Table 1. Theparticle size distribution was 5.5 μm (D10), 7.4 μm (mean particle sizeD50), 9.4 μm (D90), which were determined by using the method measuringparticle size distributions defined in Example 4. In addition, thevolume of the vehicle solution can be adjusted to prepare a formulationof the active ingredient in any concentration besides 200 mg/mL (50,100, 150, 250, 300, 400, 500, 600 mg/mL, etc.) [Example G1a].

Examples G2 to G6

The methanol used in the solution of the active ingredient in Example G1was replaced with any other organic solvent such as acetone [ExampleG2], N,N-dimethylacetamide [Example G3], 1-propanol [Example G4],2-propanol [Example G5], and dimethylsulfoxide [Example G6]) and thenthe substantially same process as Example G1 was done to prepare asustained release formulation for injection which contains 200 mg/mLcubic-shaped crystal of the active ingredient. The solvent used in eachexample and the particle size distribution of the resulting formulationswhich was measured according to the method of Example 4 are shown inTable 19.

In addition, the volume of the vehicle solution can be adjusted toprepare a formulation of the active ingredient in any concentrationbesides 200 mg/mL (50, 100, 150, 250, 300, 400, 500, 600 mg/mL, etc.)(Table 20).

TABLE 19 Organic solvent in the solution of the active ingredientConcentration of the active Particle size distribution Particle sizedistribution Particle size distribution Example No. (concentration: 70%)ingredient (mg/mL) (D10)/μm (D50; mean particle size)/μm (D90)/μm G1methanol 200 5.5 7.4 9.4 G2 acetone 200 8.4 15.1 21.4 G3 N,N- 200 4.18.8 18.9 dimethyl- acetamide G4 1- 200 10.5 18.4 25.9 propanol G5 2- 2004.8 10.3 25.2 propanol G6 dimethyl- 200 5.3 10.8 16.9 sulfoxide

TABLE 20 Organic solvent in the solution of the active ingredientConcentration of the active Example No. (concentration: 70%) ingredient(mg/mL) G1a methanol 50, 100, 150, 250, 300, 400, 500, 600, etc. G2aacetone 50, 100, 150, 250, 300, 400, 500, 600, etc. G3aN,N-dimethylacetamide 50, 100, 150, 250, 300, 400, 500, 600, etc. G4a1-propanol 50, 100, 150, 250, 300, 400, 500, 600, etc. G5a 2-propanol50, 100, 150, 250, 300, 400, 500, 600, etc. G6a dimethylsulfoxide 50,100, 150, 250, 300, 400, 500, 600, etc.

Example 12: Effect of Mean Particle Size (1) (Measurement of ParticleSize Distribution)

Table 21 shows the formulation composition of Formulations No. 4 to No.6, wherein all of the formulations have 200 mg/mL of the activeingredient.

Table 22 shows the obtained particle size distributions (D10, D50, D90)of Formulations No. 4 to 6 which were determined by the measurementmethod of particle size distribution described in Example 4. The meanparticle sizes were 5.0 μm (Formulation No. 4), 11.3 μm (Formulation No.5), and 25.8 μm (Formulation No. 6).

TABLE 21 Formulation composition Active ingredient 200 mg (free form ofCompound 1) Polysorbate 80 1.7 mg Sodium dihydrogenphosphate dihydrate1.2 mg Disodium hydrogenphosphate anhydride 2.2 mg Sodium chloride 5.8mg Water for injection 1 mL (total volume)

TABLE 22 Particle size distribution (μm) Formulation Mean particle sizeExample No. D10 D50 D90 6 4 0.5 5.0 7.9 7 5 6.3 11.3 17.5 8 6 17.5 25.835.0

(Animal Experiments)

Each of the formulations having 200 mg/mL of the active ingredient (i.e.Formulations No. 4 to No. 6) was administered to gastrocnemius muscle ofsix rats [Sprague-Dawley rats (SD), male, 9 weeks old, 250-300 g] in adose of 50 mg/kg (0.25 mL/kg). The blood was collected 1, 2, and 4hours, 1, 2, 4, 7, 14, 21, and 28 days after the administration, and theresults showed that the blood level of the present compound held upwell. As shown in FIG. 2, in case that the mean particle size was 5.0 μm(Formulation No. 4), the present compound rapidly disappeared from theblood after 7 days (1 week); whereas in case that the mean particle sizewas 11.3 μm (Formulation No. 5) and 25.8 μm (Formulation No. 6), theblood level of the present compound held up well for at least 14 days (2weeks).

Example 13: Effect of Mean Particle Size (2)

Table 23 shows the formulation composition of Formulations No. 7 to No.10, wherein all of the formulations have 400 mg/mL of the activeingredient.

Table 24 shows the obtained particle size distributions (D10, D50, D90)of Formulations No. 7 to 10 which were determined by the measurementmethod of particle size distribution described in Example 4. The meanparticle sizes were 4.8 μm (Formulation No. 7), 11.1 μm (Formulation No.8), 20.0 μm (Formulation No. 9), and 11.7 μm (Formulation No. 10).

TABLE 23 Formulation composition Active ingredient 400 mg (free form ofCompound 1) Polysorbate 80 1.3 mg Sodium dihydrogenphosphate dihydrate0.9 mg Disodium hydrogenphosphate anhydride 1.7 mg Sodium chloride 4.6mg Water for injection 1 mL (total volume)

TABLE 24 Particle size distribution (μm) Formulation Mean particle sizeExample No. D10 D50 D90 6 7 0.4 4.8 7.8 7 8 5.9 11.1 17.5 9 9 13.6 20.028.2 10 10 0.5 11.7 34.1

(Animal Experiments)

Each of the formulations having 400 mg/mL of the active ingredient (i.e.Formulations No. 7 to No. 10) was administered to gastrocnemius muscleof six rats [Sprague-Dawley rats (SD), male, 9 weeks old, 250-300 g] ina dose of 100 mg/Kg (0.25 mL/kg). The blood was collected 1, 2, and 4hours, 1, 2, 4, 7, 14, 21, and 28 days after the administration, and theresults showed that the blood level of the present compound held upwell. As shown in FIG. 3, in case that the mean particle size was 4.8 μm(Formulation No. 7), the present compound rapidly disappeared from theblood after 7 days (1 week); whereas in case that the mean particle sizewas 11.1 μm (Formulation No. 8), 20.0 μm (Formulation No. 9) and 11.7 μm(Formulation No. 10), the blood level of the present compound held upwell for at least 14 days (2 weeks).

Example 14: Effect of Mean Particle Size on Needle Passability (1)

Each of the active ingredient crystals having the following meanparticle size:

11.1 μm (Formulation No. 1, prepared in Example 1);15.2 μm (Formulation No. 2, prepared in Example 2);19.9 μm (Formulation No. 3, prepared in Example 3); and54.3 μm (Formulation No. 11, prepared in Example 11) was formulated tothree types of formulations having a concentration of 200, 300, and 400mg/mL. 1 mL of each formulation was put into a 2.5 mL syringe equippedwith a 22 gauge needle (the length of the needle=1½, Terumo). As amaterial for evaluating the needle passability, pork ham and melamineform (AISEN) were selected because these materials are similar tomuscles in the point that they require considerable power to insert theneedle into the materials. Each formulation was injected into thematerials, and then the needle passability was evaluated by judgingwhether the whole amount of each formulation could be injected or not(i.e. whether the needle's inside is clogged with the crystal or not).The results of pork ham and melamine form are shown in Table 25 andTable 26, respectively. In Table 25 and Table 26, “∘” shows that it waspossible to inject the whole amount of the formulation into thematerials, whereas “x” shows that it was difficult to inject all ofthem. As a result, there is no difference between pork ham and melamineform when they are used as a material for evaluating the needlepassability, and the results also indicate that it would be moredifficult to make an injection with the needle when the activeingredients are more concentrated and the mean particle sizes arebigger.

TABLE 25 Injection into pork ham Concentration of active FormulationMean particle ingredient (mg/mL) No. size (μm) 200 300 400 1 11.1 ∘ ∘ ∘2 15.2 ∘ ∘ ∘ 3 19.9 ∘ ∘ x 11 54.3 ∘ x x

TABLE 26 Injection into melamine form Mean particle Concentration ofactive Formulation size ingredient (mg/mL) No. (μm) 200 300 400 1 11.1 ∘∘ ∘ 2 15.2 ∘ ∘ ∘ 3 19.9 ∘ ∘ x 11 54.3 ∘ x x

Example 15: Effect of Mean Particle Size on Needle Passability (2)

In order to evaluate the effect of the mean particle size on the needlepassability in more detail, formulations of the crystal which have aconcentration of 200-400 mg/mL with a 10 mg/mL interval were preparedwith the crystals having a mean particle size of 11.1-84.9 μm. 1 mL ofeach formulation was put into a 2.5 mL syringe equipped with an 18 to 22gauge needle (the length of the needle=1½, Terumo) or a 23 gauge needle(the length of the needle=1¼, Terumo). Each formulation was injectedinto the melamine form used in Example 14, and the needle passabilitywas evaluated by judging whether the whole amount of each formulationcould be injected or not (i.e. whether the needle's inside is cloggedwith the crystal or not). Table 27 shows the results of these. In Table27, “∘” shows that it is possible to inject the whole amount of theformulation into the melamine form, whereas “x” shows that it isdifficult to inject them, and also the gauge needle is shown inbrackets. The results are as follows:

(i) when the mean particle size of the crystal was less than 54.3 μm,the formulations at a concentration of less than 220 mg/mL were able topass through a needle of 23 gauge;(ii) when the mean particle size of the crystal was less than 25.2 μm,the formulations at a concentration of less than 260 mg/mL were able topass through at least a needle of 22 gauge;(iii) when the mean particle size of the crystal was less than 19.9 μm,the formulations at a concentration of less than 300 mg/mL were able topass through at least a needle of 22 gauge;(iv) when the mean particle size of the crystal was less than 17.6 μm,the formulations at a concentration of less than 330 mg/mL were able topass through at least a needle of 22 gauge; and(v) when the mean particle size of the crystal was less than 15.2 μm,the formulations at a concentration of less than 400 mg/mL were able topass through at least a needle of 22 gauge.

TABLE 27 Mean particle Concentrations of active ingredient (mg/mL) size50- 200- 230- 270- 310- 340- (μm) 190 220 260 300 330 400 11.1 ∘ ∘ ∘ ∘ ∘∘ (18- (18- (18- (18- (18- (18- 23G) 23G) 23G) 23G) 23G) 23G) 15.2 ∘ ∘ ∘∘ ∘ ∘ (18- (18- (18- (18- (18- (18- 23G) 23G) 23G) 23G) 23G) 22G) 17.6 ∘∘ ∘ ∘ ∘ x (18- (18- (18- (18- (18- (18G) 23G) 23G) 23G) 23G) 22G) 19.9 ∘∘ ∘ ∘ x x (18- (18- (18- (18- (18G) (18G) 23G) 23G) 23G) 22G) 25.2 ∘ ∘ ∘x x x (18- (18- (18- (18G) (18G) (18G) 23G) 23G) 22G) 54.3 ∘ ∘ x x x x(18- (18- (18G) (18G) (18G) (18G) 23G) 23G) 56.1 x x x x x x (18G) (18G)(18G) (18G) (18G) (18G) 84.9 x x x x x x (18G) (18G) (18G) (18G) (18G)(18G)

INDUSTRIAL APPLICABILITY

The present invention can get more patients effectively treated becausethe present compound in the present formulation can be continuouslyreleased for at least more than 2 to 4 weeks, and the present compoundcan be also formulated into a sustained release formulation forinjection which has a good needle passability.

1. (canceled) 2: A sustained release formulation, comprising: acomposition includingN-[4-[4-(1,2-benzisothiazol-3-yl)-1-piperazinyl]-(2R,3R)-2,3-tetra-methylene-butyl]-(1′R,2′S,3′R,4′S)-2,3-bicyclo[2,2,1]heptanedicarboximideas an active ingredient, which is in a form of a crystal having a meanparticle size range within from 4 μm to 26 μm, and a surfactantcomprising at least one ingredient selected from the group consisting ofpolysorbate 80, polysorbate 20, polyoxyethylene hydrogenated castor oil50, polyoxyethylene hydrogenated castor oil 60, poloxamer 188 andpolyoxyethylene castor oil. 3: The sustained release formulation ofclaim 2, wherein the mean particle size range is 4 μm-20 μm, 4 μm-18 μm,4 μm-16 μm, 5 μm-26 μm, 5 μm-20 μm, 5 μm-18 μm, 5 μm-16 μm, 8 μm-20 μm,10 μm-26 μm, 10 μm-20 μm, 10 μm-18 μm, or 10 μm-16 μm. 4: The sustainedrelease formulation of claim 2, wherein the crystal is a cubic crystal.5: The sustained release formulation of claim 4, wherein the cubiccrystal has a length to width ratio of 1:1, and a length to height ratioof 1:0.8 to 1:1.2. 6: The sustained release formulation of claim 2,wherein the active ingredient in the form of the crystal is included in5% (w/v) to 60% (w/v) with respect to a total amount of the composition.7: The sustained release formulation of claim 2, wherein the surfactantis polysorbate
 80. 8: The sustained release formulation of claim 2,wherein the surfactant is included in a concentration of 0.05 mg/ml to20 mg/ml. 9: The sustained release formulation of claim 2, wherein thecomposition further comprises a buffer. 10: The sustained releaseformulation of claim 9, wherein the buffer is at least one ingredientselected from the group consisting of sodium phosphate, disodiumhydrogenphosphate, sodium dihydrogenphosphate, potassium phosphate,dipotassium hydrogenphosphate, potassium dihydrogenphosphate,trometamol, sodium carbonate, sodium bicarbonate, meglumine, arginine,triethanolamine and citric acid. 11: The sustained release formulationof claim 9, wherein the buffer is included in 0.01% (w/v) to 2% (w/v)with respect to a total amount of the composition. 12: The sustainedrelease formulation of claim 2, wherein the composition furthercomprises an isotonic agent. 13: The sustained release formulation ofclaim 12, wherein the isotonic agent is at least one ingredient selectedfrom the group consisting of sodium chloride and D-mannitol. 14: Thesustained release formulation of claim 12, wherein the isotonic agent isincluded in 0.1% (w/v) to 10% (w/v) with respect to a total amount ofthe composition. 15: The sustained release formulation of claim 2,wherein the composition further comprises water. 16: The sustainedrelease formulation of claim 2, wherein the active ingredient isincluded in a concentration of 50 mg/mL to 600 mg/mL. 17: The sustainedrelease formulation of claim 9, wherein the buffer is included in aconcentration of 0.1 mg/mL to 20 mg/mL. 18: The sustained releaseformulation of claim 12, wherein the isotonic agent is included in aconcentration of 1 mg/mL to 100 mg/mL. 19: The sustained releaseformulation of claim 2, which comprises 50 mg to 1200 mg of the activeingredient per a container. 20: The sustained release formulation ofclaim 2, which passes through a needle of 18 to 23 gauges. 21: Thesustained release formulation of claim 2, which is a depot formulation.22: A method for treating psychiatric disease, comprising: administeringthe sustained release formulation of claim
 2. 23: The method of claim22, wherein the psychiatric disease is schizophrenia. 24: The method ofclaim 22, wherein the psychiatric disease is bipolar disorder. 25: Themethod of claim 22, wherein the psychiatric disease is depression.